🕒 *Approximate reading time: 2 minutes*

✍️ *Author: Dr Eleni Christoforidou*

I am excited to announce that I have been featured in Beyond Science Magazine's 60th issue, where I had the opportunity to discuss my journey as a neuroscientist, my current research on amyotrophic lateral sclerosis (ALS), and my passion for science communication.

The interview touches upon various aspects of my life and career, such as:

The driving force behind my decision to become a neuroscientist
My perspective on the complexity of the brain and its implications for research
An overview of my ongoing research on ALS, its molecular mechanisms, and potential therapeutic targets
The story of how I started my science communication journey and its impact on young students
Future challenges and opportunities in the field of neuroscience
The experience of sharing my story and passion for neuroscience has been incredibly rewarding. I believe that making scientific knowledge accessible to everyone is vital in fostering curiosity, inspiring the next generation of scientists, and promoting a broader understanding of the world around us. Science communication is an essential aspect of bridging the gap between research and public perception.

As a neuroscientist, I am continuously motivated by the potential to contribute to the development of new diagnostic tools and treatments for neurological and psychiatric disorders. I am grateful for the opportunity to share my experiences with the readers of Beyond Science Magazine and hope that my story will inspire others to pursue their passion for science and make a meaningful difference in the world.

To read the full interview, please visit Beyond Science Magazine's Instagram page [here](https://www.instagram.com/p/CqVwVBJI3EY/?utm_source=ig_web_copy_link). I am looking forward to continuing my journey in neuroscience research and science communication, and I hope you'll join me in exploring the fascinating world of the human brain.

Happy reading!

![{833x1047} Beyond Sc!ence Magazine Cover, Issue 60, Special Mini Edition.](/media/BeyondScienceMagazineCover.png "Beyond Science Magazine Cover.")

I am excited to announce that I have been featured in Beyond Science Magazine's 60th issue, where I had the opportunity to discuss my...

Beyond Sc!ence Magazine Feature

🕒 *Approximate reading time: 3 minutes*

✍️ *Author: Dr Eleni Christoforidou*

In the realm of scientific research, the need for improvisation is a given. In today's lab adventure, we delve into the world of 3D printing, an innovative technology that helped standardise a cell migration assay.

![The process of designing and making 3D-printed lab equipment.](/media/3Dprint.mp4 "The process of designing and making 3D-printed lab equipment")

*Timelapse: The process of designing and making 3D-printed lab equipment.*

Our challenge today was tracking the position of individual cells over time in a migration assay, after inhibiting a protein of some immortalised cells. The goal was to ascertain if they migrated from one spot in the dish to another.

However, the scale posed a challenge. The dish was 100 mm in diameter, with each cell measuring approximately 4 μm, and over 20 million cells in the dish! The question was, how could we ensure that we were observing the exact same group of cells every time we placed the dish under the microscope?

The solution came in the form of a 3D-printed piece of plastic of specific dimensions that fit snugly beneath the dish. The design of the plastic was such that it covered the entire bottom of the dish, apart from five strategically placed holes. These holes aligned perfectly with the diameter of the field of view when looking down the microscope, allowing us to observe through these holes each time we needed to check the cells, ensuring a consistent spot over time.

This simple solution exemplifies how innovation, combined with some geometric formulae and basic programming, can significantly enhance scientific experiments.

I'm eager to hear your thoughts! Do you have any suggestions for improving this technique?

For those interested in the technical side of things, I've made the code for this 3D print available on my GitHub page [here](https://github.com/eleni-chr/100mm_dish_scratch_assay_rim).

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

Join me in the lab as we innovate with 3D-printing technology to design custom equipment that aids in the standardisation of cell migration assays in our experiments.

A Day in the Lab: Improvising with 3D-Printing for Standardised Experiments

🕒 *Approximate reading time: 3 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Today, our lab adventure takes us on a journey through the fascinating world of cells, leveraging the power of phase-contrast microscopy.

![{1746x1036} Using a phase-contrast microscope.](/media/135958.jpg "Using a phase-contrast microscope.")

Phase-contrast microscopy is a groundbreaking optical microscopy technique that transforms phase shifts in light passing through a transparent specimen into brightness variations in the resultant image. Although phase shifts are not visible in themselves, they become noticeable when displayed as alterations in brightness.

When light waves traverse a medium other than a vacuum, they interact with this medium, causing changes in wave amplitude and phase, dependent on the properties of the medium. Changes in amplitude, or brightness, stem from the scattering and absorption of light, which is often wavelength-dependent, and may give rise to colours. However, without specific provisions, phase changes remain invisible since photographic equipment and the human eye are primarily sensitive to amplitude variations.

Phase-contrast microscopy brings to light numerous cellular structures that remain hidden with a conventional bright-field microscope. Previously, these structures could only be made visible through staining, a process requiring additional preparation and the inevitable death of the cells. The advent of the phase-contrast microscope enabled biologists to study living cells in real-time, observing their proliferation through cell division. It remains one of the few methods that allow for the quantification of cellular structure and components without resorting to fluorescence.

This technique's significance and potential were so groundbreaking that its inventor, Frits Zernike, was awarded the Nobel Prize in Physics in 1953, not long after its inception in the early 1930s. Join us next time as we continue to delve into the world of cells, unearthing more of their secrets through the lens of our powerful microscopes.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

Join me in the lab as we use the groundbreaking technique of phase-contrast microscopy, a Nobel-prize winning method, to study living cells in depth and gain insights into cellular structure and proliferation.

A Day in the Lab: Harnessing a Nobel-Winning Technique

🕒 *Approximate reading time: 2 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Today, our lab adventure involves muscle biopsies, specifically those from the feet. After undergoing an immunostaining process with fluorescent antibodies, these minute biopsies are mounted on microscope slides. The aim of this process is to highlight the neuromuscular junctions, where a neuron interacts with a muscle, instructing it to either contract or relax.

![Mounting muscles on microscope slides.](/media/muscles.mp4 "Mounting muscles on microscope slides")

*Timelapse: Mounting muscle biopsies onto microscope slides.*

Post-immunostaining, these muscle samples are situated onto microscope slides, preparing them for examination using confocal microscopy. The purpose of this analysis is to ascertain whether a genetic anomaly in the motor neurons impacts the development of these neuromuscular junctions.

The image below offers an example of how these samples appear when scrutinised under the microscope.

![The neuromuscular junctions are shown in red and the neurons are shown in green. {625x624}](/media/muscles.jpg "Neuromuscular junctions under the microscope")

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

Join me today in the lab as we explore muscle biopsies under the microscope. After immunostaining them, we mount these samples on slides for in-depth scrutiny, specifically examining the intricacies of neuromuscular junctions.

A Day in the Lab: Microscopic Exploration of Muscle Biopsies

🕒 *Approximate reading time: 2 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Today's experiment in the lab involved brain cells that had been kept in cell culture flasks and nurtured in an incubator with a red solution known as the "culture medium". These flasks hosted millions of cells derived from multiple brains.

![{4032x3024} A tower of cell culture flasks at the end of my experiment.](/media/IMG_6603.jpg "Cell culture flask tower.")

A key component of the culture medium is phenol red, a pH indicator. Phenol red assumes different colours based on the pH of the environment. The picture above shows the culture medium appearing pink, a result of the pH turning excessively alkaline. This occurred as the flasks were left outside the incubator for an extended period during the experiment. Being exposed to atmospheric gases resulted in this shift in pH, as it differed from the carefully regulated environment of the incubator. If, on the other hand, the pH had leaned towards acidity, perhaps due to cells producing a surplus of waste products, the solution would have transitioned to an orange shade, eventually turning yellow.

In this colourful experiment, we learned how we could visually gauge the health of cell cultures using a pH indicator. It's another testament to how visual cues in the lab can provide vital insights into the unseen biochemical processes.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

Join me in the lab today as we delve into a colourful world of pH indicators, using phenol red to gauge pH levels in our experiment involving cultured brain cells. Let's explore the colour spectrum of pH!

A Day in the Lab: Deciphering pH Levels with Colour

🕒 *Approximate reading time: 2 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Today's lab activity involved preparing a stock of a protein for long-term storage. The process began with the introduction of a specific volume of an appropriate liquid - in this case, water - to the glass vial containing the lyophilised form of the protein. Using a syringe, the liquid was added to convert the powder into a solution of the desired concentration.

Subsequently, this solution was aliquoted, or divided, into smaller tubes. One of these aliquots was immediately used for a biochemical assay, while the remaining ones were stored at -80°C for future use.

![{1485x1037} Preparing a stock of a protein.](/media/140503.jpg "Preparing a stock of a protein")

To ensure that the protein retains its original biological or functional behaviour over the storage period - which could range from days to years - it's crucial to prevent any possible inactivation processes. The shelf life of a protein relies not only on its inherent nature but also on the storage conditions.

Proteins must be stored within an appropriate temperature and pH range, typically in the presence of a concentrated substance such as glycerol or sucrose. This helps to maintain activity and prevent protein aggregation, thereby allowing the protein to remain effective for future applications.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

Join me in the lab today as I prepare a stock of a protein for long-term storage, ensuring its integrity and functionality for future research.

A Day in the Lab: Preparing Proteins for Long-Term Storage

🕒 *Approximate reading time: 2 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Today's lab agenda revolved around the intriguing task of disrupting bacteria to extract a gene I had previously inserted into them. The tubes featured in the picture below are remnants of a bacterial culture that I had been nurturing overnight.

![{4032x3024} Bacterial culture leftovers.](/media/IMG_6796.jpg "Bacterial culture leftovers.")

Bacteria are genetic storehouses that hold more than just their chromosomal DNA. They also contain a kind of DNA known as a plasmid. To achieve my goals, I genetically engineered a plasmid to carry a specific human gene sequence. My objective was to replicate this human gene rapidly in large quantities. To accomplish this, I employed a method known as "plasmid preparation". Bacteria are prolific reproducers, and thus they duplicate their plasmids multiple times, enabling the rapid production of considerable amounts of the human gene overnight. Following this, I lysed the bacteria - meaning I used chemicals to rupture them open. This released the plasmids, which I then purified and utilised in subsequent experiments.

Today's session once again highlighted the remarkable utility of bacteria in genetic research and the fascinating methods we use to manipulate them to our advantage. These small organisms, as simple as they may seem, continue to play a significant role in advancing our understanding of genetic material and its applications.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

Join me in today's lab session where I am exploiting the power of bacteria to swiftly duplicate a human gene. Genetic amplification has never been this fascinating!

A Day in the Lab: Rapidly Amplifying Genes with Bacteria

🕒 *Approximate reading time: 3 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Today's lab adventure revolved around using a virus as a genetic courier, carrying a specific gene into a mammalian cell. This involved preparing a special disinfectant solution to neutralise the virus after the experiment. Even though the virus has been genetically modified to disable its ability to replicate and spread to other cells after infecting a host, extreme caution is exercised to ensure its proper disposal. This includes soaking all used tubes, pipette tips, gloves, and other consumables in the disinfectant before incineration.

![{1742x1037} Making up a disinfectant solution.](/media/135848.jpg "Making up a disinfectant solution")

The core of my experiment is grounded in a technique called viral transduction. This is a process in which a foreign DNA is introduced into a cell through a viral vector. Viruses, thanks to their evolutionary proficiency, have developed specialised molecular mechanisms to transport their genomes efficiently into the cells they invade. Viral vectors come with several beneficial properties:

**Safety:** The viral vectors are modified to minimise risks during handling. This usually involves removing a section of the viral genome crucial for viral replication.

**Low toxicity:** The viral vector should exert a minimal effect on the physiological condition of the infected cell.

**Stability:** Some viruses exhibit genetic instability and can rearrange their genomes rapidly. This is counterproductive to the predictability and reproducibility of work involving a viral vector, and is circumvented in their design.

**Cell type specificity:** Many viral vectors are engineered to infect a broad spectrum of cell types. However, at times the viral receptor can be modified to direct the virus towards a specific cell type.

**Identification:** Viral vectors are usually equipped with particular genes, known as markers, which help determine which cells have incorporated the viral genes. A common marker is resistance to a certain antibiotic. These cells can then be easily isolated because cells that have not absorbed the viral vector genes will lack antibiotic resistance and therefore cannot survive in its presence.

Today's session showcased the intricate interplay between virology and genetics, and the important role that viruses can play in advancing our understanding and manipulation of genetic material.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

Join me in today's lab session where I leverage a virus to transport a specific gene into a mammalian cell. A fascinating mix of genetics and virology!

A Day in the Lab: Employing Viruses to Transfer Genes into Cells

🕒 *Approximate reading time: 2 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Today's lab chronicles revolved around a chilling task - using liquid nitrogen to snap-freeze brain tissue at a brisk -196°C for long-term preservation. Dealing with liquid nitrogen required me to don appropriate Personal Protective Equipment (PPE), as showcased in the picture below.

![{1951x1951} What I looked like with all the PPE for using liquid nitrogen.](/media/6169EEDDE21C.jpg "PPE for liquid nitrogen")

PPE for liquid nitrogen usage includes cold-insulating gloves, a lab coat, non-porous closed-toe shoes, and a face shield specifically designed to guard against liquid droplets and gas. Liquid nitrogen contact with the skin can lead to severe cryogenic burns, akin to frostbite. Furthermore, liquid nitrogen should be avoided in confined and inadequately ventilated spaces because it could displace oxygen and create a suffocating environment.

An important safety note: improperly stored samples in liquid nitrogen can result in EXPLOSIONS! While I have been fortunate not to experience this personally, I was once in proximity to another scientist who had just retrieved a small tube from a liquid nitrogen tank when it suddenly exploded in his hand, causing him temporary hearing loss. This incident was likely due to trapped liquid nitrogen in the tightly sealed tube. Hence, one should never fully seal a container holding liquid nitrogen! Conversely, leaving a container with liquid nitrogen uncovered is also risky. This is because oxygen's boiling point is higher than that of nitrogen, allowing oxygen to condense from the air into the liquid nitrogen. This process converts oxygen gas from the air into liquid oxygen, which can accumulate to levels potentially causing violent and highly explosive reactions with organic materials, even normally non-flammable ones.

Liquid nitrogen is a potent tool in laboratory research, especially for preserving samples. However, safe handling is critical to prevent injuries and accidents. Always remember: proper protection, ventilation, and careful storage are the keys to working safely with liquid nitrogen.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

Let's delve into the laboratory today where I will share my experience with liquid nitrogen and how to handle it safely for long-term preservation of brain tissue.

A Day in the Lab: Safely Handling Liquid Nitrogen

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Today's lab adventure was focused on the art of pipetting. Specifically, I utilised a multichannel micropipette to expedite a biochemical assay. This allowed me to simultaneously transfer solutions to eight wells on a plate. Then, for a bit of fun and education, I snapped the photo below demonstrating the absolute worst way to handle a multichannel micropipette during your experiments. To help you perfect your pipetting technique, I've also included a list of common errors and how to avoid them.

![{2316x3088} How NOT to hold a multichannel micropipette.](/media/IMG_4670.jpg "Multichannel pipette")

ANGLE: Maintain the pipettor in an upright position. Pipetting at an angle greater than 20º affects the internal pressure in the pipette tip, compromising accuracy as the angle steepens.

DEPTH: Avoid immersing the tip too deep into the liquid as it can cause droplets to form on the tip's side or create pressure changes that modify the volume. Conversely, shallow immersion can lead to air entering the tip.

SEAL: Ensure a good seal between the pipette tip and the pipettor. A poor seal can drastically impact volumes.

SPEED: Aspirating too quickly can introduce inaccuracies, especially with viscous liquids, and potentially lead to air bubbles or liquid contact with the pipette filter, raising the risk of contamination. Dispensing liquids too quickly can cause splatter or disrupt cells at the bottom of a plate.

CONTACT: Avoid touching the tips to the bottom of a well or tube while pipetting, as this can lead to incorrect liquid volumes due to tip opening blockage. Contact with the tips can damage cell health and impact imaging and reader performance by puncturing filter membranes or damaging optical surfaces on plates.

TEMPERATURE: Remember that most pipettors depend on air pressure to move liquids, and this air pressure is influenced by temperature changes. Therefore, prolonged use can cause heat transfer from your hand to the pipette, affecting the air between the piston and liquid, and subsequently making liquid transfers less accurate the longer the pipette is used.

Using a multichannel pipette is a fantastic way to increase efficiency in the lab. However, it's essential to master the technique to ensure accuracy and maintain the integrity of your samples. Happy pipetting!

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

Join me in the lab today as I share tips on how to speed up biochemical assays using a multichannel micropipette, and common pipetting errors to avoid.

A Day in the Lab: Speeding up Pipetting

🕒 *Approximate reading time: 2 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Today's lab activity centred around a delicate and intricate task - dissecting a brain. The brain in question was preserved in a red-pink solution known as the 'dissection medium.' This solution is composed of essential nutrients designed to maintain the health and vitality of the tissue's cells during dissection until they can be transferred to an incubator for further growth. Furthermore, the medium contains a buffer that ensures the pH remains at a physiological level.

![{1744x1035} Brain dissection.](/media/140922.jpg "Brain dissection")

It's vital to perform the dissection aseptically to minimise the risk of contamination from bacteria or other potential contaminants. The dissection focused on a specific brain area, which, once isolated, was further dissected into small pieces. These fragments were then subjected to enzymatic and mechanical methods to separate the tissue into individual cells that can be cultured in an incubator.

The process of dissecting a brain is a precise and careful procedure, requiring not only technical skills but also a keen understanding of the brain's complex structures. The isolated cells will provide valuable insights for our ongoing research into brain function and pathology.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

Join me in the lab today as I delve into a brain dissection, preserving cells in a special medium for future growth and experimentation.

A Day in the Lab: Dissecting a Brain

🕒 *Approximate reading time: 3 minutes*

✍️ *Author: Dr Eleni Christoforidou*

I recently had the honor of being featured in an interview with Applied Biological Materials (abm), a company dedicated to empowering the scientific community. Our engaging conversation covered a wide range of topics, from my journey to neuroscience and experiences in grad school, to my current research and my dedication to science communication.

As a kid, my scientific interests varied widely - chemistry, nuclear physics, materials engineering, and even astronautics. But as I delved into biology during high school, I found my true passion. I was captivated by the complexities of life, which ultimately led me to neuroscience, a field I had hardly heard of until I began university applications.

In grad school, I was fortunate enough to experience a "rotation year," working in three different labs with three different supervisors. This approach enabled me to explore various research methodologies and areas, aiding in my decision to focus on neuroinflammation in neurodegenerative diseases.

Beyond research, I took our conversation to a more public platform, discussing my Instagram channel, @neuroscientist.at.work. Through this channel, I aim to make research more accessible and inspire young minds by showcasing the daily life of a scientist and grad student.

We also delved into my current research, focusing on neuroinflammation and axonal transport proteins. Despite the challenges, especially those associated with working with microglia, I remain positive and hopeful for future breakthroughs.

A typical day in the lab is diverse, ranging from cell culture and molecular biology experiments to dissections. However, I emphasised that the practical work forms less than 50% of my time. The majority is dedicated to reading scientific papers, writing protocols, doing data analysis, and learning new techniques.

The interview also touched upon my achievements, extracurricular activities, and my approach to staying organised amidst a busy schedule. I shared some advice for budding neuroscientists, emphasising the value of gaining practical experience, online courses, and seeking advice from those already in the field.

Check out the interview [here](https://info.abmgood.com/neuroscientist-at-work).

![Cool graphic created by abm to represent me and my work {1080x1080}](/media/abm.jpg)

Join me in recounting my recent SciComm Feature interview with Applied Biological Materials (abm). We discuss everything from my journey to neuroscience, my experiences in grad school, my current research, and my commitment to science communication.

My SciComm Feature with Applied Biological Materials

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Today's lab work centred on a biochemical assay to determine the concentration of a protein released by brain cells post their pro-inflammatory activation by a bacterial component. To achieve this, I administered the appropriate compounds to the cells in culture and let them incubate overnight. Subsequently, I collected the compounds released by the cells and utilised them in a sandwich ELISA (Enzyme-Linked Immunosorbent Assay). This process helps detect if the protein of interest is among the substances released by the cells and quantifies its concentration, providing insights into the cells' pro-inflammatory state post-treatment.

![Adding the samples to the ELISA plate.](/media/Elisa.mp4 "Adding the samples to the ELISA plate")

*Timelapse: Carefully loading the samples into a plate pre-coated with the capture antibody.*

## Understanding the Sandwich ELISA

A sandwich ELISA is a biochemical assay used to identify the presence of an antigen, usually a protein, in a liquid sample. This identification is achieved with the use of antibodies specifically directed against the protein under consideration. Here are the steps we follow in a sandwich ELISA:

1. Preparation of a plate by binding it with a known quantity of capture antibody.
2. Blocking any nonspecific binding sites on the surface.
3. Addition of the antigen-containing sample to the plate, where it's captured by the antibody.
4. Washing the plate to remove any unbound antigen.
5. Addition of a specific antibody that binds to the antigen, creating a 'sandwich' with the antigen trapped between two antibodies.
6. Application of enzyme-linked secondary antibodies that also bind specifically to the antibody, acting as detection antibodies. This enzyme is an amplifier, producing numerous signal molecules even if only a few enzyme-linked antibodies remain bound.
7. Another wash of the plate to remove unbound antibody-enzyme conjugates.
8. Addition of a chemical to be converted by the enzyme into a colour signal.

The resulting colour intensity is proportional to the concentration of the protein, and it can be measured by absorption spectroscopy to determine the presence and quantity of the antigen.

![Colour change before and after addition of an acid. The colour intensity is proportional to the concentration of the protein. {1135x426}](/media/ELISA.png "Colour change before and after addition of an acid")

Such biochemical assays are integral in the study of cellular behaviours and responses, providing valuable insights into various cellular processes. 

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

Dive into the world of biochemical assays with me, as I work on determining the concentration of a protein released by brain cells.

A Day in the Lab: Determining Protein Concentrations with a Biochemical Assay

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Today, the lab was abuzz with genotyping activity. We had DNA samples from the latest litters of the lab's colonies, and their genotypes needed to be determined to identify any genetic mutations in specific genes. Our techniques of choice were the Polymerase Chain Reaction (PCR) and agarose gel electrophoresis, which are routine yet integral processes in our lab, performed nearly every week. Today, I genotyped 22 DNA samples, guided by a relatively straightforward method detailed below.

![Adding loading dye](/media/Genotyping.mp4 "Adding loading dye")

_Timelapse: Adding a purple loading dye to the samples to enhance the visibility of DNA progress on the gel. The dye also contains a reagent to increase the density of the samples relative to the running buffer, ensuring the samples sink in the well._

## An In-depth Look at Today's Experiment

Genotyping determines the differences in an individual's genotype by comparing their DNA sequence for a specific gene against a reference sequence. We designed specific primers to bind to and amplify the gene of interest in the genomic DNA of a sample. This amplified gene is then run on an agarose gel - a process known as gel electrophoresis - to visualise the DNA and ascertain whether it is a wild-type or a mutant gene. Below, you can see some of the genotyping results we achieved.

![Some of the genotyping results showing the gene of interest, compared to known gene sequences and to a molecular-weight size marker (DNA ladder on the left and right ends). {555x284}](/media/14.03.19_2.jpg "Genotyping results")

These experiments provide crucial insights into the genetic profiles of our colonies, helping us understand the genetic variations at play.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

Join me in the lab as I perform genotyping to determine genetic mutations in specific genes from the latest litters of our lab's colonies.

A Day in the Lab: Determining Genotypes with PCR and Gel Electrophoresis

🕒 *Approximate reading time: 3 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Dedicated to identifying which DNA samples contained a particular mutation, I spent today in the lab conducting genotyping. The initial step involved a PCR reaction to generate DNA fragments related to our gene of interest. This was followed by gel electrophoresis, a technique that aids in visualising these fragments, subsequently identifying the samples with the desired genotype.

![Gel electrophoresis](/media/gelelectrophoresis.mp4 "Gel electrophoresis.")

*Timelapse: Carefully loading the DNA samples into a gel tank for gel electrophoresis.*

## Unravelling Gel Electrophoresis

Gel electrophoresis is a fascinating technique that separates DNA, RNA, and protein molecules based on their size and charge. It involves the movement of charged particles within an electrical current.

A gel (in the video above, I used an agarose gel I prepared earlier) acts as an anticonvective medium during electrophoresis, suppressing thermal convection caused by the application of the electric current.

The electric field consists of a negative charge at one end, which propels the molecules through the gel, and a positive charge at the other end that pulls the molecules. DNA, bearing a negative charge, is repelled by the negative charge and attracted towards the positive charge.

Shorter molecules traverse faster and migrate farther than longer ones because they navigate more easily through the pores of the gel.

To gauge their size, the DNA samples are run alongside a DNA ladder containing fragments of different known sizes.

![Gel electrophoresis in progress. Each tiny well was loaded with a DNA sample previously amplified by PCR. The purple bands are an indication of how far down the gel the samples have run. {1333x1000}](/media/gel.jpg "Gel electrophoresis in progress")

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*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

Join me as I conduct genotyping in the lab, utilising gel electrophoresis to identify DNA samples harboring a specific mutation.

A Day in the Lab: Visualising DNA through Gel Electrophoresis

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

I recently had the opportunity to sit down with The Association of Biomedical Research for an illuminating interview about my journey in the field of neuroscience. It was an enriching experience to reflect on my path, my current work, and my aspirations for the future.

In the interview, I shared insights into my research at the University of Sussex, where I am pursuing a PhD, focusing on the mechanisms of neuroinflammation in neurodegenerative diseases. The Association was particularly interested in understanding what drew me to the field and the dynamics of working in a research lab.

We delved into my career goals, which oscillate between the ambitious dream of opening my own research institute and the realistic aspiration of being a full-time researcher. I also had the chance to discuss the thrill of current projects in our lab, especially the one focusing on identifying potential biomarkers for amyotrophic lateral sclerosis.

In one of the lighter moments, I found myself reminding our audience that despite popular belief, scientists don't have all the answers! There are many everyday phenomena and diseases that still mystify us, underscoring the fact that the journey of scientific discovery is unending.

The interview was a valuable opportunity for me to articulate my experiences, aspirations, and thoughts about the intriguing world of scientific research. I hope that sharing my story can inspire others who are interested in pursuing a career in this exciting field.

Check out the interview [here](https://www.aobmr.com/post/meet-eleni-christoforidou-a-ph-d-student-at-the-university-of-sussex).

[![ABmR interview 2020](/media/abmr.png)](https://www.aobmr.org/post/meet-eleni-christoforidou-a-ph-d-student-at-the-university-of-sussex)


Join me as I recount my recent interview experience with The Association of Biomedical Research. In the blog post, I delve into the discussion about my journey in neuroscience research, my aspirations, and the captivating world of scientific discovery.

My interview with The Association of Biomedical Research

🕒 *Approximate reading time: 3 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Invested in the intricacies of brain dissection, I devoted my day in the lab to the delicate procedure of removing the meninges. This task is a vital precursor to the harvesting of certain brain sections required for glial cell culture. Purging the meninges ensures that our cell cultures remain unadulterated by meningeal fibroblasts.

![Removing the meninges](/media/Removingmeninges.mp4 "Removing the meninges.")

*Timelapse: Delicately removing the meninges from a tiny brain.*

## Shedding Light on the Meninges

Functioning as a protective envelope, the meninges comprise three membranes known as the dura mater, the arachnoid mater, and the pia mater, collectively safeguarding the brain and spinal cord. Their primary purpose is to shield the central nervous system from any external intrusions, thus ensuring its integrity and functionality.

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*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

Dive into the meticulous procedure of brain dissection, focusing on the careful removal of the meninges, a crucial step for effective glial cell culture.

A Day in the Lab: Removing the Meninges from the Brain

🕒 *Approximate reading time: 3 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Dedication and meticulousness were the essence of today's lab work, where I found myself entirely immersed in cryopreserving brain cells for future use. An elaborate process, it consumed roughly three hours, uninterrupted, to diligently prepare a modest batch of 10 samples.

![Cryopreservation](/media/Cryopreservation.mp4 "Cryopreservation.")

*Timelapse: The detailed process of cryopreserving cells.*

## Unveiling the Intricacies of Cryopreservation

One might wonder about the safeguarding measures taken to prevent these delicate cells from perishing due to freezing during the process. The answer lies in a specific cryoprotectant known as dimethyl sulfoxide (DMSO). This compound plays a crucial role in the cryopreservation process as it helps curtail the formation of damaging ice crystals, which otherwise would be fatal to the cells.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

Explore the intricate process of cryopreserving brain cells in the lab, ensuring their viability for future experimentation.

A Day in the Lab: Cryopreservation of Brain Cells

🕒 *Approximate reading time: 3 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Today in the lab, I was deeply engrossed in a miniprep experiment, an intriguing process that facilitates the purification of plasmid DNA from bacterial cultures. The previous day, I meticulously prepared overnight cultures of bacteria which I had previously transformed using a vector plasmid. This plasmid incorporates a specific human gene which is fundamental to my upcoming experiments. As a result of their overnight growth, the bacteria multiplied profusely, consequently producing an abundance of gene copies. Today, my primary objective was to extract the plasmid DNA housing the gene from these bacterial cultures.

![Miniprep](/media/Miniprep.mp4 "Miniprep")

_Timelapse: Diving into the preparation of bacterial samples for the extraction of plasmid DNA._

## A Deep Dive into Today's Experimental Methodology

The day kicked off with bacterial lysis under alkaline conditions, leading to the denaturation of chromosomal DNA and proteins. However, the plasmid DNA, with the human gene embedded in it, remained stable throughout. This method is fondly referred to as a 'miniprep' owing to its quick and small-scale nature in isolating plasmid DNA. The yield from each bacterial culture typically ranges between 50-100 μg of plasmid DNA. An absolute wonder of modern molecular biology.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post unveils the fascinating procedure of a miniprep experiment, aiming to purify plasmid DNA from bacterial cultures.

A Day in the Lab: Extracting Plasmid DNA from Bacterial Cultures

🕒 *Approximate reading time: 3 minutes*

✍️ *Author: Dr Eleni Christoforidou*

In the lab today, I embarked on the meticulous task of dissecting neonatal brains. The goal was to extract glial cells and culture them for my ongoing experiments. The task was time-intensive; dissecting a total of seven brains occupied roughly 6.5 hours of my day, all without pause. The procedure itself is reasonably straightforward, albeit requiring exceptional dexterity given the delicate and diminutive nature of the neonatal brains.

![{1090x1000} Neonatal brains meticulously dissected for today's riveting experiment.](/media/img_20191220_120342.jpg "Brains")

## Delving Deeper into Today's Experimental Methodology

From each brain, the hippocampi and cortices were diligently dissected and minced using spring scissors. The tissue harvested from each brain was then transferred into a cold dissection medium, ready for the addition of trypsin to dissociate the cells.

Following this, the cells were incubated in a 37°C water bath for 15 minutes, ensuring frequent swirling. Next, a trypsin inhibitor was introduced to halt trypsin activity, with the tube left to incubate for a brief period (1 minute) at room temperature.

Subsequently, DNase I was added to the mix to digest the sticky DNA released from the dead cells, followed by centrifugation of the tube at 400 × g for 5 minutes. Once the supernatant was removed, the pellet was triturated in pre-warmed culture medium, then subjected to another round of centrifugation at the same speed.

Finally, after the supernatant's removal, the pellet was resuspended in 5 ml of pre-warmed culture medium. This resulting cell suspension was then cultured in a PDL-coated T-75 tissue culture flask, again with pre-warmed culture medium, and incubated under specific conditions (37°C, 5% CO2).

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

In this blog post, I delve into the intricate process of dissecting neonatal brains to extract glial cells, followed by culturing them for further scientific experiments.

A Day in the Lab: Delving into Neonatal Brain Dissection for Glial Cells

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

In the ever-evolving field of neuroscience, the CRISPR/Cas9 gene-editing system has been nothing short of revolutionary. This innovative tool allows scientists to make precise alterations to DNA sequences, and thus offers significant potential for understanding and treating neurological disorders.

## Understanding the Basics of CRISPR/Cas9

CRISPR/Cas9, which stands for Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated protein 9, is a system adapted from a naturally occurring genome editing system in bacteria.

Here’s a simplified explanation of how it works:

The CRISPR sequence is a short RNA sequence that matches the DNA sequence to be edited.

The Cas9 enzyme acts like a pair of molecular scissors, cutting the DNA at the location specified by the CRISPR sequence.

The cell’s natural DNA repair machinery recognises the cut and repairs the DNA, either by sticking the ends back together, or by inserting a new piece of DNA at the cut site.

## CRISPR/Cas9 in Neuroscience

CRISPR/Cas9 has many potential applications in neuroscience:

1. Understanding Neurological Disorders: By creating animal models with specific genetic mutations associated with neurological disorders, scientists can study the disease mechanisms in a controlled setting.

2. Identifying Therapeutic Targets: Gene-editing with CRISPR/Cas9 can help identify the function of specific genes and their products, potentially uncovering novel therapeutic targets for drug development.

3. Developing Gene Therapies: In theory, CRISPR/Cas9 could be used to correct disease-causing mutations in the brains of people with genetic neurological disorders.

## CRISPR/Cas9 and Neurodegenerative Diseases

Gene editing using the CRISPR/Cas9 system has already been instrumental in advancing our understanding of neurodegenerative diseases:

1. Modelling Diseases: Scientists have used CRISPR/Cas9 to create animal models of diseases like Alzheimer’s, Parkinson’s, and ALS, providing valuable insights into these diseases.

2. Unveiling Disease Mechanisms: For example, scientists have used CRISPR/Cas9 to understand how mutations in the C9orf72 gene cause ALS and frontotemporal dementia.

3. Potential Therapeutic Application: Preliminary studies suggest CRISPR/Cas9 could be used to reduce the levels of disease-causing proteins in mouse models of neurodegenerative diseases.

## Conclusion

CRISPR/Cas9 is a powerful tool that is revolutionising our approach to studying and potentially treating neurological disorders. While there are still technical and ethical challenges to overcome before it can be widely applied in a clinical setting, the potential of this gene-editing technology in neuroscience is truly exciting.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post delves into the transformative role of the CRISPR/Cas9 gene-editing system in neuroscience, exploring its potential in understanding neurological disorders, identifying therapeutic targets, and developing innovative gene therapies.

CRISPR/Cas9: Revolutionising gene editing in neuroscience

🕒 *Approximate reading time: 3 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Today's exploration in the lab led me into the intricate realm of brain microglia. These cells were selectively isolated from a mixed culture containing both microglia and astrocytes, a task accomplished three days prior. These microglia, originally dissected from neonatal brains, were then cultured in a flask, priming them for today's investigation.

To examine them, I utilised a phase-contrast microscope, offering me a clear and detailed perspective of their microscopic world. I was particularly interested in analysing the morphology of the microglia and the purity of the culture, essential factors to confirm the success of the isolation experiment. The image below unveils the captivating sight of the cells I inspected today.

![{1333x1000} Intricately cultured microglia, extracted from a mixed glia culture of cells originally dissected out of neonatal brains.](/media/img_7948_microglia_20x_06.11.2019.jpg "Microglia")

## Diving Deeper into Today's Experimental Approach

The isolated microglia were cultivated in the lab using a gentle trypsinisation method. Initially, the culture medium was withdrawn, and the cells were rinsed with Dulbecco's Phosphate-Buffered Saline (DPBS). The mixed glia-conditioned medium underwent a 0.2 μm filter treatment to eliminate any cell debris.

The cells were then treated with trypsin-EDTA, diluted 1:4 in Dulbecco’s Modified Eagle’s Medium (DMEM), and incubated under specific conditions (37°C, 5% CO2) for 45 minutes. This step served to detach the astrocyte layer, and the trypsin activity was subsequently halted by adding culture media.

Following this, the medium was fully aspirated to discard the detached astrocytes and oligodendrocytes, and the cells were thoroughly washed with DPBS. The cells left behind were the adhered microglia, which were then nurtured in 50% mixed glia-conditioned culture medium under optimal conditions (37°C, 5% CO2) for two days until they resumed their ramified form.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post offers an inside look at the investigation of brain microglia, dissected from neonatal brains and cultured in a lab, scrutinising their morphology and culture purity using a phase-contrast microscope.

A Day in the Lab: Investigating Isolated Microglia from Neonatal Brains

🕒 *Approximate reading time: 3 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Today, in the lab, my task was twofold: to verify the health of cells thriving in culture and to estimate their population. The confluency of these cells - the degree to which they cover the culture surface - is of prime importance as it informs whether they are ready to take part in upcoming experiments.

One indispensable tool in my daily tissue culture examinations is the phase-contrast microscope. This ingenious instrument allows me to peer into the microscopic universe of these cells, ensuring their growth aligns with expectations.

The beauty of the phase-contrast microscope lies in its capacity to convert subtle phase shifts in light, caused when it traverses through transparent cells, into discernible changes in brightness that construct an image. Although these phase shifts are invisible to the naked eye, they burst into visibility as variations in brightness under the microscope. To achieve this transformation, the microscope expertly separates the illuminating (background) light from the light scattered by the specimen, constituting the foreground details.

A fascinating outcome of this process is that the cells appear to glow - a positive indicator of their health and vitality, as demonstrated in the image below.

![Phase-contrast microscopy](/media/phase-contrast.mp4 "Phase-contrast microscopy.")

*Timelapse: Ensuring the robust health and anticipated growth of my glial cells under the watchful gaze of the phase-contrast microscope.*

![Image from a phase-contrast microscope, capturing microglial cells. As these cells spend more time in culture, they begin to ramify, developing more and more processes. {1333x1000}](/media/IMG_7988.JPG "Cells under a phase-contrast microscope")

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post takes you on a journey into the lab, showcasing how phase-contrast microscopy is employed to inspect the health and confluency of cultured cells, a critical factor before they are utilised in scientific experiments.

A Day in the Lab: The Art and Science of Phase-contrast microscopy

🕒 *Approximate reading time: 3 minutes*

✍️ *Author: Dr Eleni Christoforidou*

On this day in the lab, I embarked on an exciting journey into the microscopic world of bacterial transformation. My task? To facilitate a unique experiment which allows E. coli bacteria to clone themselves, thereby producing multiple copies of a gene that has piqued my scientific curiosity.

The goal of my experiment was to modify these ubiquitous E. coli bacteria by introducing a fragment of DNA known as a vector. This vector harbours the gene for a green fluorescent protein (GFP), which will serve as a crucial element in my upcoming investigations. Intriguingly, the vector also carries a gene for antibiotic resistance, a key feature for the success of this endeavour.

Here's why: The agar plates, where the bacteria are left to proliferate overnight, contain an antibiotic that prohibits the growth of unwanted bacterial visitors. Only the bacteria bearing the antibiotic resistance gene - which, importantly, are also the carriers of our GFP gene - can thrive on these plates.

![Bacterial transformation](/media/Bacterial-Transformation.mp4 "Bacterial transformation.")

_Timelapse: Distributing the bacteria on agar plates for an overnight incubation session, resulting in a thriving colony of cloned bacteria._

## Unfolding the Techniques Behind Today's Experiment

Bacterial transformation, the process employed today, represents one of three routes for horizontal gene transfer. This fascinating process enables a bacterial cell to uptake exogenous DNA. For successful transformation, the bacteria must be in a state of competence, which can be artificially induced in the lab.

Our procedure involves exposing the bacteria to calcium chloride under cool conditions, followed by a swift heat shock. This combined assault disrupts the cell membrane partially, creating an opportunity for the DNA to be ushered into the cell. This is where the science of genetic modification takes centre stage, setting the stage for a new chapter in our understanding of these microscopic marvels.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post presents a snapshot of a day in the lab spent conducting a bacterial transformation experiment, resulting in genetically modified bacteria cloning and expressing a specific gene of interest.

A Day in the Lab: Bacterial transformation

🕒 *Approximate reading time: 2 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Today's activities in the lab were centred around inoculating four flasks of bacterial cultures. These flasks will be incubated overnight to allow the bacteria to clone themselves and replicate a piece of DNA that was introduced into them. The bacteria are cultured in a nutrient-rich medium known as lysogeny broth, composed of peptides, vitamins, minerals, and other components to ensure optimal bacterial growth.

![Bacterial cultures](/media/Bacterial-Cultures.mp4 "Bacterial cultures.")

*Timelapse: The video showcases the preparation of flasks with lysogeny broth, followed by their inoculation with bacteria.*

## Delving Deeper into the Timelapse Video

The glass barrier visible between myself and the samples is part of a laminar flow cabinet, a vital piece of equipment in microbiology labs designed to prevent sample contamination. The cabinet operates by drawing air through a High-Efficiency Particulate Air (HEPA) filter and projecting it in a laminar flow towards the user. A laminar flow implies that the air flows smoothly in layers with high momentum diffusion and low momentum convection, resulting in minimal mixing (contrary to turbulent flow). This direction of airflow safeguards the sample from the user, though it does not protect the user from the sample.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

A day in the lab, dedicated to inoculating bacterial cultures to enable self-cloning and DNA replication. Discover the process, from broth preparation to sterile conditions.

A Day in the Lab: Inoculating bacterial cultures

🕒 *Approximate reading time: 3 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Today's activities in the lab were directed at the in-depth analysis of calcium activity in the white matter located within the corpus callosum, the region of the brain bridging the two hemispheres. The process engaged recordings of the brain obtained through a 2-photon microscope. A red fluorescent dye enabled the visualisation of blood vessels, while a green fluorescent dye highlighted the white matter (essentially, the oligodendrocytes encircling the neurons). The purpose of this study was to scrutinise the spontaneous calcium activity within the white matter, a venture that was unprecedented. This task necessitated significant coding via Matlab and a host of complex mathematical functions to extract pertinent information from the microscopic recordings.

Here's an example of what the actual recordings appeared like under the microscope.

![Two-photon microscope image](/media/8.mp4 "Two-photon microscope image")
_Timelapse: An illustration of a 2-photon imaging session, showcasing blood vessels in red, and brain neurons in green. You can observe the individual red blood cells journeying through the vessels as dark shadows. The rise in green signal indicates the calcium activity of the oligodendrocytes (or the white matter), the insulating material around the neurons._

## An Insight into Today's Experimental Procedures

The experiment involved the employment of a genetically-encoded calcium indicator in oligodendrocytes, facilitating the recording of calcium activity in myelin in vivo. This was achieved using 2-photon imaging, via a cranial window fitted over the corpus callosum. Computer code was then employed to identify individual oligodendrocytes and extract their basal calcium activity. This data was then compared under different conditions (3 weeks post-surgery, 5 weeks post-surgery, during locomotion, and at rest). An array of mathematical functions, such as Principal Component Analysis (PCA) and Independent Component Analysis (ICA), were utilised for data analysis. These functions were crucial for extracting cellular signals from the imaging datasets and for computing the firing rate of the cells.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

Take a peek into a day at the lab, focused on exploring the calcium activity of the white matter within the corpus callosum, utilising 2-photon microscopy, intricate coding, and innovative methodologies.

A Day in The Lab: Investigating Calcium Activity in the White Matter

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Since their discovery in 2006, induced pluripotent stem cells (iPSCs) have revolutionised the field of regenerative medicine and disease modelling. In this blog post, we explore how iPSCs are being utilised in the research of neurodegenerative diseases.

## What are Induced Pluripotent Stem Cells?

iPSCs are a type of stem cell that can be generated directly from adult cells. Through the introduction of specific genes, scientists can reprogramme somatic cells, like skin or blood cells, into iPSCs. These cells maintain the defining properties of stem cells: the ability to self-renew and to differentiate into various cell types, including neurons.

## iPSCs and Neurodegenerative Disease Research

Neurodegenerative diseases, such as Alzheimer's and Parkinson's, involve the progressive loss of neurons and are often characterised by the accumulation of abnormal proteins. They are notoriously difficult to study, as obtaining live neurons from patients for research purposes is not feasible. This is where iPSCs come into play.

Scientists can reprogramme cells from a patient with a neurodegenerative disease into iPSCs. These cells can then be differentiated into neurons, creating a personalised in vitro model of the patient's nervous system. This provides an unprecedented opportunity to study the pathophysiology of these diseases and test potential treatments.

## The Advancements and Challenges

Using iPSCs, scientists have been able to model various neurodegenerative diseases and gain significant insights into their molecular mechanisms. For example, iPSC-derived neurons from patients with Alzheimer's have been shown to produce amyloid-beta, a protein that forms the hallmark plaques seen in the brains of these patients.

However, as with any rapidly evolving field, challenges exist. For instance, iPSC-derived neurons are typically in a developmental state, which might not perfectly mirror the aged neurons typically affected in neurodegenerative diseases. In addition, iPSC technology requires significant expertise and resources, limiting its accessibility.

## Conclusion

Despite these challenges, the use of iPSCs in neurodegenerative disease research remains an exciting and rapidly evolving field. As technology advances, iPSCs will continue to provide valuable insights into these diseases and accelerate the development of effective treatments.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post explores the use of induced pluripotent stem cells (iPSCs) in neurodegenerative disease research, highlighting their potential to revolutionise our understanding and treatment of these challenging conditions.

The Use of Induced Pluripotent Stem Cells in Neurodegenerative Disease Research

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

The world of medicine continues to make incredible strides, and stem cell research is at the forefront of many of these advancements. Stem cells, with their ability to self-renew and differentiate into multiple cell types, are emerging as a promising tool in treating various conditions, including neurodegenerative diseases.

## What Are Stem Cells?

Stem cells are unique cells with the capacity to develop into any cell type in the body. They can divide and create identical copies of themselves (a process called self-renewal) or differentiate into specialised cells. This dual potential makes stem cells invaluable in the world of regenerative medicine.

Stem##  Cells and Neurodegenerative Diseases

Neurodegenerative diseases, including Alzheimer's, Parkinson's, and ALS, are characterised by progressive loss of structure or function of neurons. Despite extensive research, there are currently no definitive cures for these diseases, only treatments to manage symptoms. However, stem cell therapy offers hope for new therapeutic approaches.

Stem cells could potentially be used to replace damaged neurons in these diseases. In theory, stem cells could be introduced into the brain where they would differentiate into neurons and other cell types, replacing those that have been lost to disease.

Research into this approach is ongoing, but early results from preclinical studies and clinical trials have been promising. For example, trials using stem cells to treat Parkinson's disease have shown evidence of improved motor function in some patients.

## Challenges and Future Perspectives

Despite the promise, there are still many challenges to overcome. For instance, ensuring the newly introduced stem cells differentiate into the correct cell type and integrate properly with the existing neural networks. Additionally, we must ensure the procedure is safe and doesn't introduce new health risks, such as the potential for tumour formation.

Yet, with continual advancements in our understanding and ability to manipulate stem cells, we are steadily overcoming these challenges. The future of stem cell therapy in treating neurodegenerative diseases looks promising, holding hope for millions of people worldwide.

## Conclusion

Stem cell therapy is a burgeoning field that holds immense potential for treating neurodegenerative diseases. While challenges remain, the future is promising, and with each passing day, we move a step closer to turning this potential into reality.

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*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post delves into the potential of stem cell therapy as a promising approach to treat neurodegenerative diseases, discussing current research, challenges, and future perspectives.

The Potential of Stem Cell Therapy in Treating Neurodegenerative Diseases

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

For many years, the brain was thought to be a relatively static organ after a certain stage of development. It was believed that once an individual reached adulthood, their brain was largely fixed in its structure and function. However, more recent research has revealed that the brain is in fact a highly dynamic and adaptable organ, capable of significant change throughout a person's lifetime. This remarkable ability is known as neuroplasticity.

## Understanding Neuroplasticity

Neuroplasticity refers to the ability of the brain to change its structure and function in response to experience. This can include creating new neurons (a process known as neurogenesis), forming new connections between neurons (synaptic plasticity), and even rerouting or transforming existing connections.

## Implications of Neuroplasticity

The implications of neuroplasticity are vast and wide-ranging. For one, it suggests that our brains are not set in stone, but rather can be reshaped and reorganised by our experiences. This has significant implications for learning and memory, suggesting that the more we use certain neural pathways, the stronger they become.

Neuroplasticity also has profound implications for recovery from brain injury. It suggests that with the right stimulation and rehabilitation, the brain can reorganise itself to compensate for damage, potentially restoring lost functions. This is a promising area of research for conditions such as stroke and traumatic brain injury.

## Challenges and Future Directions

While the concept of neuroplasticity is exciting, it's important to note that our understanding of this phenomenon is still developing. We don't yet fully understand how neuroplasticity works, or how to harness it effectively for therapeutic purposes.

However, ongoing research is continually shedding light on this fascinating aspect of brain function, and holds promise for the development of new interventions to enhance learning, memory, and recovery from brain injury.

## Conclusion

Neuroplasticity, the brain's remarkable ability to adapt and change, challenges many traditional beliefs about the brain. As we continue to explore this fascinating aspect of neuroscience, we are discovering new ways in which our brains are shaped by our experiences, and uncovering exciting possibilities for enhancing brain health and function.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post explores the concept of neuroplasticity, the brain's remarkable ability to change its structure and function in response to experience, and its implications for learning, memory, and recovery from brain injury.

Neuroplasticity: The Brain's Ability to Adapt and Change

🕒 *Approximate reading time: 5 minutes*

✍️ *Author: Dr Eleni Christoforidou*

The past few decades have seen remarkable advancements in our understanding of neurological disorders, but translating this knowledge into effective treatments remains a significant challenge. One promising area of research is the application of nanotechnology to the diagnosis, monitoring, and treatment of these disorders.

## Defining Nanotechnology

Nanotechnology is a multidisciplinary field that manipulates matter at the nanoscale (1-100 nanometres) to create new materials and devices. These nanoscale entities have unique properties that can be leveraged for various applications in medicine, including targeted drug delivery, improved imaging, and regenerative medicine.

## Nanotechnology for Targeted Drug Delivery

One of the most promising applications of nanotechnology in neurology is targeted drug delivery. Neurological disorders often require therapeutic agents to be delivered directly to the brain, but many drugs cannot cross the blood-brain barrier (BBB), a protective barrier that prevents potentially harmful substances in the blood from entering the brain. Nanotechnology can help overcome this obstacle. For instance, nanoparticles can be engineered to carry drugs across the BBB, enabling targeted delivery of therapeutic agents to specific brain regions or cell types.

## Nanotechnology in Neuroimaging

In addition to drug delivery, nanotechnology holds potential for improving neuroimaging. Nanoparticles can be used as contrast agents in magnetic resonance imaging (MRI), enhancing the visibility of specific tissues or cellular processes. This could enable earlier and more accurate diagnosis of neurological conditions and better monitoring of disease progression and treatment response.

## Nanotechnology and Regenerative Medicine

Nanotechnology also has potential in the field of regenerative medicine. For instance, nanomaterials can be used to create scaffolds for tissue engineering, providing a supportive structure for the growth of new neurons or glial cells. This could be particularly useful in conditions such as spinal cord injury or stroke, where significant neuronal loss occurs.

## Conclusion

While nanotechnology holds exciting potential for treating neurological disorders, significant challenges remain, such as ensuring the safety and biocompatibility of nanomaterials, and refining techniques for targeted drug delivery and imaging. Nevertheless, as research in this field progresses, it’s likely that nanotechnology will become an increasingly important tool in our fight against neurological disorders.

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*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post discusses the promising role of nanotechnology in the diagnosis and treatment of neurological disorders, highlighting its applications in targeted drug delivery, neuroimaging, and regenerative medicine.

The Potential of Nanotechnology in Treating Neurological Disorders

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Oligodendrocytes are a type of glial cell found in the central nervous system (CNS). These cells are vital for neuronal health, predominantly due to their role in producing myelin, a fatty substance that insulates neuronal axons, enhancing the speed and efficiency of electrical signal transmission.

## Meet the Oligodendrocytes

Oligodendrocytes are the CNS equivalent of the peripheral nervous system's Schwann cells. Both types of cells produce myelin, but while a single Schwann cell myelinates just one segment of one peripheral nerve fibre, a single oligodendrocyte can extend its processes to 40-50 axon segments, thereby myelinating multiple axon segments simultaneously.

## The Myelin Sheath: An Essential Insulator

Myelin is composed of various lipids and proteins and forms a protective sheath around axons. This sheath not only protects axons from damage but also facilitates the rapid propagation of electrical signals, or action potentials, along the axon. Without myelin, these signals would travel significantly slower, impairing neural communication.

## Oligodendrocytes, Myelin, and Neurological Disorders

Given the importance of oligodendrocytes and myelin in neuronal function, it's not surprising that damage to these cells or to the myelin sheath can lead to serious neurological conditions. Multiple sclerosis, for instance, is characterised by the degradation of the myelin sheath, leading to slower neural communication, muscle weakness, and other debilitating symptoms.

## Promoting Oligodendrocyte and Myelin Health

Research into the preservation and repair of oligodendrocytes and the myelin sheath is a hot topic in neuroscience. Some promising avenues include the investigation of specific growth factors that encourage oligodendrocyte proliferation, and exploration of drugs that could protect myelin from damage.

## Conclusion

Oligodendrocytes and myelin are crucial components of the CNS, playing pivotal roles in maintaining neuronal function. Understanding how these components can be protected and restored could have significant implications for the treatment of a range of debilitating neurological disorders.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post discusses the roles of oligodendrocytes and the myelin they produce in the central nervous system, explaining their importance in neuronal function and the implications for neurological disorders when these components are damaged.

Oligodendrocytes and Myelin: Protecting and Supporting Neuronal Function

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

The gut-brain axis is an intriguing concept that has captivated researchers' interest in recent years. It encompasses the bi-directional communication between the gut and the brain, and it is becoming clear that the state of our gut health can significantly impact our brain function.

## The Gut-Brain Axis: A Two-Way Street

The gut and brain are intimately connected, with the gut often referred to as the 'second brain'. This is due to the gut’s complex neural network and its extensive communication with the brain. These interactions take place via the autonomic nervous system, the enteric nervous system, the immune system, and through the circulatory system.

## The Role of the Microbiome

The gut microbiome, the community of trillions of bacteria living in our digestive tract, plays a vital role in this communication. The microbiota can produce neurotransmitters and other metabolites that can influence brain function and behaviour. Dysbiosis, or an imbalance in the gut microbiota, has been linked to various neurological and psychiatric disorders, including anxiety, depression, autism, and Parkinson's disease.

## How Does the Gut Communicate with the Brain?

The gut communicates with the brain via several mechanisms:

1. Vagus Nerve: The vagus nerve, the longest cranial nerve, provides the primary route for gut-brain communication. Stimulation or inhibition of the vagus nerve can have significant effects on mood and cognitive function.

2. Immune System: The gut microbiota can influence the immune system, and thereby indirectly affect the brain.

3. Metabolites and Hormones: The gut microbiota produces a range of metabolites, such as short-chain fatty acids, and hormones that can influence brain function.

## Promoting Gut Health for Better Brain Health

Maintaining a healthy gut microbiome is therefore crucial for brain health. This can be achieved through a balanced diet rich in fibre, regular exercise, adequate sleep, and reducing stress levels.

## Conclusion

The gut-brain axis represents an exciting area of research with potential therapeutic implications for various neurological and mental health disorders. However, more research is needed to fully understand this complex relationship and to develop effective therapeutic strategies.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post delves into the fascinating concept of the gut-brain axis, exploring how the gut and its resident microbiota communicate with the brain and how gut health can significantly impact brain function and behaviour.

The Gut-Brain Axis: How Gut Health Affects the Brain

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Our ability to form and store memories is a complex process that involves numerous changes at the molecular level within our neurons. This blog post delves into the molecular underpinnings of memory formation and storage.

## The Neuronal Changes Underlying Memory

Memory formation involves structural changes at the synaptic level in neurons, a process known as synaptic plasticity. This includes the strengthening or weakening of synapses, which facilitates the storage of information. These changes occur through two major molecular processes: Long-Term Potentiation (LTP) and Long-Term Depression (LTD).

1. Long-Term Potentiation (LTP): This process refers to the long-lasting strengthening of synapses based on recent patterns of activity. This strengthening is believed to contribute to the formation of memories.

2. Long-Term Depression (LTD): The opposite of LTP, LTD involves the long-lasting decrease in synaptic strength, which is also critical for memory formation and learning.

## The Molecular Players in Memory Formation

Several molecular players are involved in these processes, including neurotransmitters, receptors, and intracellular signalling molecules.

1. Neurotransmitters and Receptors: Glutamate, the main excitatory neurotransmitter in the brain, binds to its receptors on the postsynaptic neuron, leading to the activation of various intracellular signalling pathways. Among these receptors, NMDA and AMPA receptors play a key role in LTP and LTD.

2. Intracellular Signalling: Activation of these receptors triggers intracellular signalling cascades involving various molecules such as calcium, protein kinases, and transcription factors. These molecules, in turn, affect gene expression and protein synthesis, leading to changes in the structure and function of the synapse.

## Consolidation and Storage of Memory

Once formed, memories undergo consolidation, a process that transforms short-term memories into long-term ones. This involves gene expression and protein synthesis, which are required for structural changes at the synapse. The precise molecular mechanisms of memory storage, however, remain a subject of ongoing research.

## Conclusion

While we have a basic understanding of the molecular basis of memory formation and storage, many questions remain. Continued research in this field promises to further unravel the complex molecular machinery that enables us to form, store, and retrieve memories.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

In this blog post, we delve into the fascinating world of the molecular processes that underpin memory formation and storage. We explore synaptic plasticity, the key molecular players involved, and the processes of memory consolidation and storage.

The Molecular Basis of Memory Formation and Storage

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Autophagy, a cellular process that ensures the degradation and recycling of damaged components, is crucial for maintaining cellular health. Interestingly, disruptions in autophagy have been linked to neurodegenerative diseases, opening up new avenues for research and potential therapies.

## What is Autophagy?

Autophagy is a cellular self-eating process where damaged proteins, organelles, and other cellular constituents are enclosed in a double-membraned vesicle known as an autophagosome. The autophagosome then fuses with a lysosome, an organelle filled with enzymes capable of breaking down the cell's waste, to form an autolysosome where the waste is degraded and recycled.

## Autophagy in Neuronal Health

Neurons are heavily dependent on autophagy due to their high metabolic activity and their inability to dilute damaged components by cell division, as they are post-mitotic cells. Autophagy aids in maintaining the health of neurons by removing damaged proteins and organelles, which could otherwise accumulate and interfere with neuronal function.

## Autophagy and Neurodegenerative Diseases

There's a growing body of evidence suggesting a connection between disrupted autophagy and neurodegenerative diseases:

1. Alzheimer's Disease: Research has identified a reduction in autophagy in the brains of patients with Alzheimer's disease, which could lead to an accumulation of misfolded proteins and neuronal death.

2. Parkinson's Disease: Mutations in genes encoding proteins involved in autophagy have been identified in familial cases of Parkinson's disease, suggesting a role for autophagy dysfunction in the disease's pathogenesis.

3. Huntington's Disease: Impaired autophagy is also observed in models of Huntington's disease, leading to the build-up of harmful protein aggregates.

4. Amyotrophic Lateral Sclerosis (ALS): Altered autophagy has been reported in both sporadic and familial forms of ALS.

## Targeting Autophagy in Neurodegenerative Diseases

Given the importance of autophagy in maintaining neuronal health and its apparent role in neurodegenerative diseases, strategies to modulate autophagy are being explored as potential treatments. For example, drugs that enhance autophagy could potentially be used to clear accumulated proteins in diseases like Alzheimer's and Parkinson's.

## Conclusion

While there is still much to understand about the complex role of autophagy in neurodegeneration, it is clear that this fundamental cellular process is a key player in neuronal health and disease. Unravelling the intricacies of autophagy and its relationship with neurodegenerative diseases holds promise for developing innovative therapeutic strategies.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post explores the intriguing relationship between autophagy, a vital cellular process, and neurodegenerative diseases. We discuss the role of autophagy in neuronal health, its link to various neurodegenerative diseases, and the potential for targeting autophagy in therapeutic strategies.

The Role of Autophagy in Neurodegeneration

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Amyotrophic lateral sclerosis (ALS) is a complex neurodegenerative disease characterised by the progressive loss of motor neurons in the brain and spinal cord. The exact causes of ALS remain elusive, but it is believed that a combination of genetic and environmental factors contribute to the disease's development and progression. Among the various cellular and molecular mechanisms implicated in ALS, cytoplasmic dynein 1 has emerged as a protein of interest due to its role in several cellular processes and potential involvement in ALS pathology.

### Cytoplasmic dynein 1: An Overview

Cytoplasmic dynein 1 is a motor protein belonging to the dynein family. It is responsible for the transport of various cellular cargoes, including vesicles, organelles, and protein complexes, along microtubules towards the cell center. This process, known as retrograde transport, is critical for maintaining cellular homeostasis and ensuring proper intracellular communication.

In addition to cargo transport, cytoplasmic dynein 1 plays a role in various other cellular processes, such as cell division, autophagy, and the positioning of cellular organelles like the Golgi apparatus and endoplasmic reticulum. Given its involvement in these essential functions, it is not surprising that dysregulation of cytoplasmic dynein 1 activity has been implicated in several neurodegenerative diseases, including ALS.

### Cytoplasmic dynein 1 and ALS

While the precise role of cytoplasmic dynein 1 in ALS is still being explored, several lines of evidence suggest its involvement in the disease's pathology:

1. Altered expression and function: Studies have shown that the expression and function of cytoplasmic dynein 1 are altered in ALS patients and animal models of the disease. These alterations can lead to impaired retrograde transport, contributing to motor neuron degeneration by disrupting cellular homeostasis and communication.

2. Interaction with ALS-associated proteins: Cytoplasmic dynein 1 has been found to interact with several proteins implicated in ALS, such as TAR DNA-binding protein 43 (TDP-43) and fused in sarcoma (FUS). The abnormal aggregation of these proteins in motor neurons is a hallmark of ALS, and their interaction with cytoplasmic dynein 1 may disrupt its normal function, further contributing to the disease's progression.

3. Genetic mutations: While mutations in the cytoplasmic dynein 1 gene are rare, some studies have identified mutations in genes encoding dynein-associated proteins, such as dynactin, in familial and sporadic ALS cases. These mutations can impair the function of the dynein-dynactin complex, leading to defects in retrograde transport and motor neuron degeneration.

### Therapeutic Implications

Given the potential involvement of cytoplasmic dynein 1 in ALS pathology, targeting this protein or its associated pathways could represent a promising therapeutic approach. Some potential strategies include:

1. Enhancing dynein function: Small molecules or other therapeutic agents that enhance dynein function or stabilise the dynein-dynactin complex could potentially improve retrograde transport and reduce motor neuron degeneration in ALS patients.

2. Modulating protein-protein interactions: Targeting the interactions between cytoplasmic dynein 1 and ALS-associated proteins, such as TDP-43 or FUS, could help prevent the disruption of dynein function and subsequent motor neuron degeneration.

3. Boosting cellular clearance mechanisms: Enhancing autophagy or other cellular clearance pathways could help remove aggregated ALS-associated proteins and restore dynein function, ultimately protecting motor neurons from degeneration.

4. Gene therapy: For ALS cases with mutations in dynein-associated genes, gene therapy approaches could be employed to correct the mutations and restore the normal function of the dynein-dynactin complex.

### Conclusion

Cytoplasmic dynein 1 is a motor protein involved in numerous essential cellular processes, and its dysfunction has been implicated in the pathology of amyotrophic lateral sclerosis (ALS). While the precise role of cytoplasmic dynein 1 in ALS is still being elucidated, targeting this protein or its associated pathways could represent a promising therapeutic strategy for this devastating neurodegenerative disease. Continued research on cytoplasmic dynein 1 and its interactions with other cellular components will be critical for advancing our understanding of ALS and developing novel treatments to slow or halt the progression of the disease.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

In this blog post, we discuss the potential role of cytoplasmic dynein 1, a motor protein involved in essential cellular processes, in the pathology of amyotrophic lateral sclerosis (ALS) and explore how targeting this protein or its associated pathways could represent a promising therapeutic strategy for the disease.

The Role of Cytoplasmic Dynein 1 in ALS Pathology

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Bioinformatics is an interdisciplinary field that combines computer science, statistics, mathematics, and biology to analyse and interpret complex biological data. In amyotrophic lateral sclerosis (ALS) research, bioinformatics has become an indispensable tool for deciphering the wealth of data generated by high-throughput technologies like genomics, transcriptomics, and proteomics, ultimately contributing to a better understanding of the disease's molecular mechanisms and informing the development of novel therapeutic strategies.

### The Role of Bioinformatics in ALS Research

Bioinformatics has played a crucial role in various aspects of ALS research, including:

1. Genome-wide association studies (GWAS): By analysing large genomic datasets, bioinformatics has helped identify risk loci and candidate genes associated with ALS, providing insights into the disease's genetic architecture and potential therapeutic targets.

2. Transcriptome analysis: Bioinformatics tools are essential for processing and interpreting the massive amounts of data generated by RNA-sequencing, enabling the identification of differentially expressed genes, alternative splicing events, and non-coding RNA species implicated in ALS pathology.

3. Network analysis: Bioinformatics approaches can be used to construct and analyse protein-protein interaction networks, metabolic networks, and gene regulatory networks, offering a systems-level perspective on the complex molecular processes underlying ALS.

### Therapeutic Implications

By helping to decipher the complex data generated by high-throughput technologies, bioinformatics can inform the development of more effective treatments for ALS:

1. Identifying drug targets: Bioinformatics can help prioritise candidate genes and pathways for therapeutic intervention based on their association with ALS and their biological relevance.

2. Drug repurposing: By comparing the molecular signatures of ALS with those of other diseases, bioinformatics can facilitate the identification of existing drugs that could be repurposed for ALS treatment.

3. Personalised medicine: Bioinformatics approaches can be used to analyse individual patient data, potentially enabling the development of personalised treatment strategies based on a patient's unique genetic and molecular profile.

### Conclusion

Bioinformatics has emerged as a critical component of amyotrophic lateral sclerosis (ALS) research, enabling researchers to analyse and interpret complex biological data and providing valuable insights into the disease's molecular mechanisms. By leveraging bioinformatics approaches, scientists can identify novel therapeutic targets, facilitate drug repurposing, and develop personalised medicine strategies, ultimately contributing to the development of more effective treatments for this devastating neurodegenerative disease.

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*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

In this blog post, we discuss the importance of bioinformatics in amyotrophic lateral sclerosis (ALS) research and how this interdisciplinary field contributes to the identification of novel therapeutic targets, drug repurposing, and personalised medicine strategies.

Using bioinformatics to unravel the mysteries of amyotrophic lateral sclerosis (ALS)

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Amyotrophic lateral sclerosis (ALS) is a complex neurodegenerative disease characterised by the progressive loss of motor neurons in the brain and spinal cord. While the precise causes of ALS remain largely unknown, various cellular and molecular mechanisms have been implicated in the disease's development and progression. One such mechanism involves the protein TAR DNA-binding protein 43 (TDP-43), which has been found to play a significant role in the pathology of ALS.

### TDP-43: An Overview

TDP-43 is a highly conserved, ubiquitously expressed nuclear protein involved in various aspects of RNA metabolism, including transcription, splicing, and transport. It also plays a role in the regulation of gene expression and the formation of stress granules, which are transient cytoplasmic aggregates that form in response to cellular stress.

### TDP-43 and ALS

The involvement of TDP-43 in ALS pathology was first discovered when it was identified as a major component of the protein aggregates found in the affected motor neurons of most ALS patients. Since then, several lines of evidence have further implicated TDP-43 in the disease:

1. Abnormal localisation and aggregation: In ALS, TDP-43 is often mislocalised from the nucleus to the cytoplasm, where it forms insoluble aggregates. These aggregates are thought to be toxic to motor neurons, contributing to their degeneration.

2. Loss of nuclear function: The mislocalisation of TDP-43 from the nucleus can lead to a loss of its normal nuclear functions, potentially disrupting RNA metabolism and contributing to motor neuron dysfunction.

3. Genetic mutations: Mutations in the TDP-43 gene (TARDBP) have been identified in both familial and sporadic ALS cases, further implicating the protein in the disease's pathogenesis.

### Therapeutic Implications

Given the involvement of TDP-43 in ALS pathology, targeting this protein or its associated pathways could represent a promising therapeutic approach. Some potential strategies include:

1. Preventing TDP-43 mislocalisation and aggregation: Small molecules or other therapeutic agents that stabilise TDP-43 in its native state or enhance its nuclear localisation could potentially reduce its aggregation and toxicity.

2. Boosting cellular clearance mechanisms: Enhancing autophagy or other cellular clearance pathways could help remove aggregated TDP-43 and restore normal protein homeostasis in motor neurons.

3. Modulating TDP-43-related RNA metabolism: Targeting the RNA metabolism pathways regulated by TDP-43 could help alleviate motor neuron dysfunction caused by the loss of TDP-43 nuclear function.

### Conclusion

TDP-43 is a crucial protein involved in various aspects of RNA metabolism, and its dysfunction has been implicated in the pathology of amyotrophic lateral sclerosis (ALS). Targeting TDP-43 or its associated pathways could represent a promising therapeutic strategy for ALS. Continued research on TDP-43 and its interactions with other cellular components will be critical for advancing our understanding of ALS and developing novel treatments to slow or halt the progression of the disease.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

In this blog post, we delve into the role of TAR DNA-binding protein 43 (TDP-43) in the pathology of amyotrophic lateral sclerosis (ALS), discussing its involvement in RNA metabolism and how targeting this protein or its associated pathways may offer promising therapeutic strategies for the disease.

TDP-43 and its Involvement in the Pathology of Amyotrophic Lateral Sclerosis (ALS)

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

RNA-sequencing (RNA-seq) is a high-throughput sequencing technology that allows researchers to study the transcriptome – the complete set of RNA transcripts produced by the genome – in a given sample. By providing a comprehensive view of gene expression, RNA-seq has become a valuable tool for investigating the molecular mechanisms underlying various diseases, including amyotrophic lateral sclerosis (ALS), a neurodegenerative disorder characterised by the progressive loss of motor neurons in the brain and spinal cord.

### The Power of RNA-sequencing

RNA-seq offers several advantages over traditional methods of gene expression analysis, such as microarrays:

1. High sensitivity and specificity: RNA-seq can detect both lowly and highly expressed transcripts, as well as discriminate between closely related genes and isoforms.

2. Unbiased analysis: Unlike microarrays, RNA-seq does not rely on predetermined probes, allowing for the unbiased discovery of novel transcripts, isoforms, and fusion genes.

3. Quantitative accuracy: RNA-seq provides precise quantification of gene expression levels, enabling the detection of subtle changes in transcript abundance.
RNA-sequencing in ALS Research

RNA-seq has been widely employed in ALS research, providing valuable insights into the disease's molecular mechanisms and identifying potential therapeutic targets:

1. Differential gene expression: RNA-seq has been used to identify genes and pathways that are differentially expressed in ALS patients compared to healthy controls, shedding light on the processes that drive motor neuron degeneration.

2. Alternative splicing: RNA-seq has revealed widespread alterations in alternative splicing – the process by which different mRNA isoforms are generated from the same gene – in ALS, highlighting its potential role in disease pathogenesis.

3. Non-coding RNA expression: RNA-seq has facilitated the discovery of novel non-coding RNAs, such as microRNAs and long non-coding RNAs, that may contribute to ALS pathology by regulating gene expression.

### Therapeutic Implications

By providing a comprehensive view of the ALS transcriptome, RNA-seq has the potential to identify novel therapeutic targets and inform the development of more effective treatments for the disease:

1. Targeting dysregulated genes and pathways: The identification of differentially expressed genes in ALS can inform the development of therapies aimed at modulating the activity of these genes or their associated pathways.

2. Splicing modulators: The discovery of widespread alterations in alternative splicing in ALS raises the possibility of using splicing modulators to correct these abnormalities and ameliorate motor neuron dysfunction.

3. Non-coding RNA-based therapies: The identification of dysregulated non-coding RNAs in ALS opens up new avenues for therapeutic intervention, such as using antisense oligonucleotides or small molecules to modulate their expression or function.

### Conclusion

RNA-sequencing has emerged as a powerful tool for investigating gene expression in amyotrophic lateral sclerosis (ALS), providing valuable insights into the disease's molecular mechanisms and identifying potential therapeutic targets. Continued research employing RNA-seq and other high-throughput technologies will be crucial for advancing our understanding of ALS and developing novel treatments to slow or halt the progression of this devastating neurodegenerative disease.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

In this blog post, we discuss the role of RNA-sequencing in amyotrophic lateral sclerosis (ALS) research, highlighting how this powerful technology has advanced our understanding of the disease's molecular mechanisms and informed the development of novel therapeutic strategies.

RNA-sequencing: A powerful tool for investigating gene expression in amyotrophic lateral sclerosis (ALS)

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Microglia, the primary immune cells of the central nervous system, play a critical role in maintaining neuronal health and responding to injury or disease. Emerging evidence suggests that microglia-derived microRNAs (miRNAs) could be involved in the pathogenesis of amyotrophic lateral sclerosis (ALS), a devastating neurodegenerative disease characterised by the progressive loss of motor neurons in the brain and spinal cord.

### MicroRNAs: Small but Powerful Regulators

MicroRNAs are small, non-coding RNA molecules that regulate gene expression post-transcriptionally. By binding to complementary sequences in target messenger RNAs (mRNAs), miRNAs can suppress translation or promote mRNA degradation, ultimately modulating the levels of specific proteins. Given their regulatory roles, dysregulation of miRNA expression or function has been implicated in various diseases, including neurodegenerative disorders like ALS.

### Microglia-derived MicroRNAs and ALS

Several lines of evidence suggest that microglia-derived miRNAs could contribute to the pathogenesis of ALS:

1. Altered miRNA expression: Studies have shown that the expression of specific miRNAs is altered in microglia in both ALS patients and animal models of the disease. These changes in miRNA expression can influence the production of various proteins involved in neuroinflammation, motor neuron survival, and other cellular processes relevant to ALS pathology.

2. Neuroinflammation: Microglia-derived miRNAs can modulate the inflammatory response in the central nervous system. In ALS, dysregulated miRNA expression may contribute to a chronic pro-inflammatory state, which can exacerbate motor neuron degeneration.

3. Neuron-microglia communication: miRNAs can be secreted by microglia and taken up by neighboring neurons, influencing their gene expression and function. In the context of ALS, this intercellular communication may contribute to the spread of disease pathology and motor neuron dysfunction.

### Therapeutic Implications

The potential involvement of microglia-derived miRNAs in ALS pathogenesis raises the possibility of targeting these molecules or their associated pathways for therapeutic intervention. Some potential strategies include:

1. Modulating miRNA expression or function: Small molecules, antisense oligonucleotides, or other therapeutic agents could be used to modulate the expression or function of specific miRNAs implicated in ALS pathology, potentially ameliorating neuroinflammation and motor neuron degeneration.

2. Enhancing neuron-microglia communication: Promoting beneficial neuron-microglia communication by modulating miRNA secretion or uptake could help counteract the detrimental effects of dysregulated miRNA expression in ALS.

3. Combination therapies: Given the complex nature of ALS, combining miRNA-based therapies with other approaches targeting different aspects of the disease may provide the most effective treatment strategy.

### Conclusion

Microglia-derived microRNAs have emerged as potential players in the pathogenesis of amyotrophic lateral sclerosis (ALS), with roles in neuroinflammation, motor neuron survival, and neuron-microglia communication. Targeting these miRNAs or their associated pathways could represent a promising therapeutic strategy for ALS. Continued research on the role of microglia-derived miRNAs in neurodegenerative diseases will be crucial for advancing our understanding of ALS and developing novel treatments to slow or halt the progression of the disease.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post explores the potential role of microglia-derived microRNAs in the pathogenesis of amyotrophic lateral sclerosis (ALS), discussing their involvement in neuroinflammation and neuron-microglia communication, and highlighting the therapeutic implications of targeting these molecules or their associated pathways to develop novel treatments for this devastating neurodegenerative disease.

Microglia-derived microRNAs: A potential player in the pathogenesis of Amyotrophic Lateral Sclerosis (ALS)

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Microscopy is an indispensable tool in neuroscience research, providing a window into the intricate world of neurons and their surrounding environment. By enabling scientists to visualise cellular structures, connections, and molecular processes at various scales, microscopy has not only advanced our understanding of the nervous system but has also revealed the beauty and complexity of the neuronal landscape.

### A Glimpse into the World of Microscopy in Neuroscience

Several types of microscopy techniques have been instrumental in neuroscience research:

1. Light microscopy: Traditional light microscopy techniques, such as bright-field and phase-contrast microscopy, allow for the visualisation of cellular morphology and the investigation of basic neuronal properties.

2. Fluorescence microscopy: Fluorescence microscopy, which includes techniques like confocal and two-photon microscopy, enables the visualisation of specific cellular structures, proteins, or molecules by utilising fluorescent labels, such as fluorescent proteins or dye-conjugated antibodies.

3. Electron microscopy: Electron microscopy, including transmission electron microscopy (TEM) and scanning electron microscopy (SEM), offers much higher resolution than light microscopy, enabling researchers to visualise subcellular structures and even individual synaptic connections.

4. Super-resolution microscopy: Techniques like stimulated emission depletion (STED) microscopy and structured illumination microscopy (SIM) overcome the diffraction limit of light microscopy, providing unprecedented resolution and allowing researchers to investigate the nanoscale organisation of neuronal structures.

### Unraveling the Mysteries of the Neuronal Landscape

Microscopy has been crucial in deciphering various aspects of neuronal biology:

1. Neuronal morphology: Microscopy techniques have allowed scientists to visualise and classify different types of neurons based on their unique shapes, dendritic branching patterns, and axonal projections.

2. Synaptic connections: Microscopy has enabled the identification and characterisation of synapses – the specialised junctions where neurons communicate – and the study of synaptic plasticity, a key mechanism underlying learning and memory.

3. Cellular processes: By visualising subcellular structures and molecular processes, microscopy has provided insights into the mechanisms governing neuronal function, including neurotransmitter release, ion channel activity, and intracellular signaling pathways.

4. Neuronal degeneration: Microscopy has been critical in characterising the pathological changes associated with neurodegenerative diseases like ALS, such as protein aggregation, mitochondrial dysfunction, and synaptic loss.

### The Beauty and Complexity of the Neuronal Landscape

Beyond its scientific utility, microscopy has also revealed the breathtaking beauty and complexity of the neuronal landscape:

1. Neuronal networks: Microscopy images have captured the intricate and highly interconnected networks formed by neurons, illustrating the remarkable complexity of the nervous system.

2. Subcellular architecture: High-resolution microscopy has unveiled the intricate organisation of neuronal compartments, such as dendritic spines and synaptic clefts, showcasing the fine balance between form and function in neurons.

3. Dynamic processes: Time-lapse microscopy has allowed researchers to visualise dynamic cellular processes, such as axonal growth, synaptogenesis, and neuronal migration, revealing the orchestrated ballet of cellular events that shape the developing nervous system.

### Conclusion

The art of microscopy has been pivotal in exploring the neuronal landscape, offering a window into the intricate world of neurons and their surrounding environment. By allowing scientists to visualise cellular structures, connections, and molecular processes at various scales, microscopy has not only advanced our understanding of the nervous system but has also unveiled the beauty and complexity of the neuronal world.

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*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post delves into the world of microscopy in neuroscience research, highlighting its critical role in unraveling the mysteries of the neuronal landscape and showcasing the breathtaking beauty and complexity of the nervous system.

The Art of Microscopy: Exploring the Neuronal Landscape

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Microglia, the primary immune cells of the brain, are integral to maintaining neural homeostasis and responding to injuries or diseases. The ability to culture these cells in vitro has greatly enhanced our understanding of their role in neurological conditions, such as amyotrophic lateral sclerosis (ALS).

## The Steps Involved in Microglia Culture

1. Tissue Dissection: Microglia are typically isolated from brain tissue, often derived from neonatal mice. The brain tissue is carefully dissected and the meninges, the layers of tissue covering the brain, are removed.

2. Enzymatic Dissociation: The tissue is then enzymatically dissociated using an enzyme like collagenase or trypsin. This process breaks down the extracellular matrix and cell-cell connections, freeing the individual cells.

3. Centrifugation and Resuspension: The dissociated cells are then centrifuged, and the cell pellet is resuspended in a suitable medium to create a cell suspension.

4. Density Gradient Separation: A density gradient centrifugation step can be included to enrich for microglia. During centrifugation, the cells separate based on their size and density, allowing for the collection of a microglia-enriched fraction.

5. Plating and Culture: The isolated microglia are plated onto tissue culture plates and incubated in a controlled environment, where they can adhere, grow, and divide.

## Studying Microglia in Culture

Once microglia are in culture, they can be studied in several ways. For instance, researchers may:

1. Analyse their Morphology: Microglia have distinct morphological states, ranging from a ramified (resting) state to an amoeboid (activated) state. Observing these changes can provide insights into their functional status.

2. Examine their Protein Expression: Immunofluorescence staining or Western blotting can be used to assess the expression of specific proteins, such as markers of activation or inflammation.

3. Monitor their Phagocytic Activity: Microglia play a crucial role in clearing debris and dead cells from the brain. Assays that track the uptake of labelled particles can help study this function in vitro.

4. Investigate their Response to Stimuli: Microglia can be exposed to various stimuli, such as pro-inflammatory cytokines or components of pathogens, to understand their response mechanisms.

## Microglia Culture in the Context of Neurodegenerative Diseases

Cultured microglia have been instrumental in studying neurodegenerative diseases like ALS:

1. Modelling Disease Mechanisms: Microglia from transgenic animals modelling ALS can help unravel disease-specific changes in microglial function.

2. Investigating Neuroinflammation: In ALS and other neurodegenerative conditions, microglia often exhibit an activated, pro-inflammatory state. Cultured microglia can help us understand the triggers and consequences of this inflammation.

3. Screening Therapeutics: Microglial cultures can also be used to screen potential therapeutics that aim to modulate microglial function or reduce neuroinflammation.

## Conclusion

The cultivation of microglia has provided researchers with a valuable tool for studying these essential brain cells. Through the continued refinement of microglial culture techniques, we are poised to gain further insights into the role of these cells in health and disease, potentially paving the way for novel therapeutic strategies in neurodegenerative diseases such as ALS.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post delves into the process of microglia culture, exploring how these resident immune cells of the brain are isolated and studied in vitro, and their significance in researching neurodegenerative diseases like ALS.

Microglia Culture: Isolating and Studying the Brain's Immune Cells

🕒 *Approximate reading time: 5 minutes*

✍️ *Author: Dr Eleni Christoforidou*

In this blog post, you will find an overview of amyotrophic lateral sclerosis (ALS), exploring its causes, risk factors, symptoms, and current treatments, while also highlighting ongoing research efforts to develop new therapies and improve our understanding of this devastating neurodegenerative disease.

### Causes and Risk Factors

The exact cause of ALS remains unknown, but it is believed to involve a combination of genetic and environmental factors. In about 5-10% of cases, the disease is inherited, and these cases are referred to as familial ALS. The remaining 90-95% of cases are sporadic, meaning they occur without a known family history. While specific genes have been identified in some cases of familial ALS, the genetic factors contributing to sporadic ALS are still largely unknown.

Some of the known risk factors for ALS include:

1. Age: ALS is most commonly diagnosed between the ages of 40 and 70, although it can occur at any age.

2. Sex: Men are slightly more likely to develop ALS than women, although this difference decreases with age.

3. Family history: Having a close relative with ALS increases the risk of developing the disease.

4. Environmental factors: Some studies suggest that exposure to certain chemicals or toxins, smoking, and a history of head injuries may increase the risk of ALS.

### Symptoms

The symptoms of ALS can vary greatly from person to person, but typically, they include:

1. Muscle weakness, particularly in the limbs, which may manifest as difficulty walking, gripping objects, or lifting the arms.

2. Muscle cramps and twitching.

3. Slurred speech or difficulty swallowing.

4. Impaired breathing as the disease progresses.

5. In some cases, cognitive and behavioral changes.

### Diagnosis

There is no single diagnostic test for ALS. The diagnosis is typically made based on clinical signs and symptoms, along with tests to rule out other conditions that may cause similar symptoms. These tests may include blood and urine tests, nerve conduction studies, electromyography (EMG), and imaging studies such as magnetic resonance imaging (MRI).

### Treatment and Management

Unfortunately, there is currently no cure for ALS. The primary goal of treatment is to manage symptoms and improve the quality of life for patients. Some strategies for managing ALS include:

1. Medications: Riluzole and edaravone are FDA-approved drugs that may slow the progression of ALS. Other medications may be used to manage specific symptoms such as muscle cramps, pain, and mood changes.

2. Physical therapy: Physical therapy can help maintain muscle strength and flexibility, improve mobility, and prevent joint stiffness.

3. Occupational therapy: Occupational therapists can recommend adaptive devices and strategies to help patients maintain their independence in daily activities.

4. Speech therapy: Speech therapists can help patients with communication difficulties and suggest alternative communication methods when necessary.

5. Breathing support: As respiratory muscles weaken, patients may require breathing assistance through non-invasive ventilation or, in some cases, invasive ventilation via a tracheostomy.

6. Nutritional support: Maintaining a healthy diet and weight is crucial for ALS patients. A dietitian can help develop a meal plan that meets the patient's nutritional needs and addresses any swallowing difficulties.

7. Emotional and psychological support: Living with ALS can be challenging, and patients may benefit from counseling or support groups to help cope with the emotional and psychological aspects of the disease.

### Research and Future Perspectives

Although there is no cure for ALS yet, researchers are continually working to better understand the underlying mechanisms of the disease and develop new therapeutic strategies. Some of the most promising areas of research include:

1. Identifying novel genetic factors and molecular pathways involved in ALS, which could lead to new therapeutic targets.

2. Investigating the role of neuroinflammation and immune cells in ALS progression.

3. Developing gene therapies and stem cell therapies to replace or repair damaged motor neurons.

4. Exploring the potential of neuroprotective agents to slow or halt the progression of the disease.

In conclusion, amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease that primarily affects motor neurons, leading to progressive muscle weakness and paralysis. While there is currently no cure for ALS, ongoing research efforts hold promise for the development of new therapies and a better understanding of the disease's underlying mechanisms. By raising awareness and supporting research, we can strive towards a future where ALS is no longer an untreatable condition.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

In this blog post, you will find an overview of amyotrophic lateral sclerosis (ALS), exploring its causes, risk factors, symptoms, and current treatments, while also highlighting ongoing research efforts to develop new therapies and improve our understanding of this devastating neurodegenerative disease.

Understanding the Basics of Amyotrophic Lateral Sclerosis (ALS)

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Cell culture techniques play a crucial role in neuroscience research, allowing scientists to study the biology of neurons and glial cells in a controlled environment. By providing a platform for the investigation of cellular processes, cell culture has contributed to numerous advances in our understanding of the nervous system and the development of novel therapeutic strategies for neurodegenerative diseases like amyotrophic lateral sclerosis (ALS).

### A Brief Overview of Cell Culture Techniques in Neuroscience

Neuroscience research relies on various types of cell culture systems, including:

1. Primary neuronal cultures: Primary neurons are isolated directly from animal tissues, such as the brain or spinal cord, and cultured in vitro. These cultures provide a physiologically relevant environment for studying neuronal development, function, and plasticity.

2. Glial cell cultures: Glial cells, such as astrocytes, oligodendrocytes, and microglia, play essential roles in supporting and regulating neuronal function. Culturing these cells allows researchers to investigate their interactions with neurons and their involvement in various neurological disorders.

3. Immortalised cell lines: Immortalised cell lines, such as neuroblastoma or glioblastoma cells, can be cultured indefinitely and provide a consistent, easily accessible source of cells for high-throughput screening and large-scale experiments.

4. Co-cultures and organotypic cultures: Co-cultures involve growing multiple cell types together, while organotypic cultures maintain the three-dimensional architecture of a specific brain region. Both systems allow researchers to investigate cell-cell interactions and the effects of the cellular microenvironment on neuronal function and survival.

### Applications of Cell Culture Techniques in Neuroscience Research

Cell culture techniques have been instrumental in advancing our understanding of the nervous system and the development of novel therapeutic strategies for neurodegenerative diseases:

1. Mechanistic studies: Cell culture systems enable researchers to dissect the molecular and cellular mechanisms underlying neuronal development, function, and dysfunction in various neurological disorders, including ALS.

2. Drug discovery: Cell culture models can be used to screen and validate potential therapeutic compounds, helping to accelerate the drug discovery process for neurological diseases.

3. Gene editing and gene therapy: With the advent of gene-editing technologies like CRISPR/Cas9, cell culture systems provide an ideal platform for studying the effects of genetic modifications on neuronal function and survival, as well as developing gene therapy approaches for neurological disorders.

4. Disease modeling: The development of induced pluripotent stem cell (iPSC) technology has revolutionised disease modeling, allowing researchers to generate patient-specific neuronal and glial cell types to study the cellular and molecular basis of neurological disorders.

### Conclusion

Cell culture techniques are of paramount importance in neuroscience research, providing a versatile platform for investigating the biology of neurons and glial cells, dissecting the mechanisms underlying neurological disorders, and developing novel therapeutic strategies for neurodegenerative diseases like ALS. As cell culture technologies continue to evolve, their applications in neuroscience research will undoubtedly continue to expand, offering new opportunities for understanding and treating complex neurological disorders.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post highlights the importance of cell culture techniques in neuroscience research, exploring their applications in mechanistic studies, drug discovery, gene editing, and disease modeling, and their contributions to our understanding of the nervous system and the development of novel therapeutic strategies for neurodegenerative diseases.

The Importance of Cell Culture Techniques in Neuroscience Research

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Neuroinflammation is a double-edged sword in the context of neurodegenerative diseases. While it is a necessary process for dealing with injury and infection, chronic neuroinflammation can exacerbate disease progression.

## Understanding Neuroinflammation

Neuroinflammation is the inflammatory response within the brain or spinal cord. This process is primarily mediated by microglia, the resident immune cells of the central nervous system (CNS). In response to injury or infection, microglia become activated and release pro-inflammatory cytokines to combat the threat. However, if this response becomes chronic, it can contribute to the damage rather than protecting the brain.

## Neuroinflammation as a Friend

When functioning properly, neuroinflammation plays a critical role in maintaining the health of the CNS. It helps in clearing pathogens, removing damaged cells, and promoting recovery and repair. Acute neuroinflammation can be seen as a protective response aimed at restoring homeostasis.

## Neuroinflammation as a Foe

However, when neuroinflammation becomes chronic, it can contribute to neurodegenerative diseases. Chronic neuroinflammation is implicated in several conditions, including Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis. It is characterised by sustained microglial activation and constant release of pro-inflammatory cytokines, leading to ongoing damage and neuronal death.

## Balancing the Neuroinflammation Response

Understanding how to balance the beneficial and harmful effects of neuroinflammation is crucial for developing new therapeutic strategies. One approach could involve dampening chronic inflammation while preserving the acute inflammatory response necessary for repair and recovery.

## Conclusion

Neuroinflammation is a complex process with both beneficial and harmful effects on the CNS. The challenge for researchers is to untangle this complexity and identify therapeutic strategies that can harness the protective aspects of neuroinflammation while minimising its damaging effects.

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*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post explores the double-edged sword of neuroinflammation in neurodegenerative diseases. We discuss its role as both a protective mechanism and a potential contributor to disease progression, and the ongoing research to balance these effects for therapeutic benefit.

Neuroinflammation: Friend or Foe in Neurodegenerative Diseases?

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Western blotting is an essential molecular biology technique frequently utilised in neuroscience research. This workhorse tool allows for the detection and quantification of specific proteins in a sample, thus aiding in the exploration of complex biological questions.

## The Basics of Western Blotting

The process of Western blotting comprises a few key stages:

1. Gel Electrophoresis: Proteins from the sample are separated according to their molecular weight by an electric field in a gel matrix.

2. Transfer: The separated proteins are then transferred from the gel to a membrane, typically made of nitrocellulose or polyvinylidene fluoride (PVDF).

3. Blocking and Probing: To prevent non-specific binding, the membrane is first blocked with a protein-rich solution. It is then probed with antibodies that recognise and bind to the protein of interest.

4. Detection: A secondary antibody, which recognises the first, is applied. This secondary antibody is linked to an enzyme that catalyses a reaction, producing a detectable signal corresponding to the protein of interest.

## Applications of Western Blotting in Neuroscience

1. Protein Expression Studies: Western blotting can reveal the levels of a specific protein in neuronal cells or tissues, enabling scientists to understand changes in protein expression under different conditions, such as disease states or after drug treatment.

2. Protein Modification Studies: Western blotting can also detect post-translational modifications of proteins, such as phosphorylation or ubiquitination, which play crucial roles in neuronal function.

3. Verification of Antibodies: Researchers can use Western blotting to verify the specificity of antibodies they plan to use in other experiments, like immunohistochemistry or immunofluorescence.

## Western Blotting in the Study of Neurodegenerative Diseases

For neurodegenerative diseases such as ALS, Western blotting is a vital technique:

1. Protein Misfolding and Aggregation: Many neurodegenerative diseases involve the misfolding and aggregation of specific proteins. Western blotting can detect these protein aggregates, providing insights into disease mechanisms.

2. Effect of Therapeutics: If a potential therapeutic is designed to increase or decrease the level of a specific protein, Western blotting can be used to confirm whether the treatment has had the desired effect.

3. Biomarker Discovery: By comparing protein levels in healthy and diseased tissues, researchers can identify potential biomarkers for disease diagnosis or progression.

## Conclusion

Western blotting is a versatile and invaluable tool in neuroscience research, providing critical insights into protein expression and modification. Despite its limitations, such as semi-quantitative nature and labour-intensity, its wide applications, from studying disease mechanisms to validating potential therapeutics, ensure its continuing role in the neuroscience research arsenal.

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*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post explores the method and applications of Western blotting in neuroscience, shedding light on its role in protein expression studies, protein modification studies, and its significant contribution to neurodegenerative disease research.

Western blotting: A versatile tool in neuroscience research

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Immunofluorescence staining is a powerful technique that allows scientists to visualise specific proteins or other molecules within cells or tissues. In neuroscience research, this method has been essential for investigating the molecular and cellular processes underlying neuronal function and dysfunction, providing invaluable insights into the pathogenesis of neurodegenerative diseases like amyotrophic lateral sclerosis (ALS).

## The process of immunofluorescence staining involves several steps:

1. Fixation: The cells or tissues are first fixed to preserve their structure and immobilise the proteins or other molecules of interest.
2. Permeabilisation: The cells or tissues are treated with a permeabilising agent, such as Triton X-100, to allow the antibodies to access intracellular targets.
3. Blocking: The samples are incubated with a blocking solution to prevent non-specific binding of the antibodies.
4. Primary antibody incubation: The samples are incubated with a primary antibody that specifically recognises the target molecule.
5. Secondary antibody incubation: The samples are incubated with a secondary antibody that recognises the primary antibody and is conjugated to a fluorescent dye.
6. Imaging: The samples are visualised under a fluorescence microscope, and the location of the target molecule is revealed by the fluorescence emitted by the dye.

## Applications of Immunofluorescence Staining in Neuroscience

Immunofluorescence staining has a wide range of applications in neuroscience research, including:

1. Cellular and subcellular localisation: Immunofluorescence staining can be used to determine the distribution of a specific protein or other molecule within a cell or tissue, providing insights into its potential functions.
2. Detection of post-translational modifications: Specific antibodies can be used to detect post-translational modifications, such as phosphorylation or ubiquitination, which can regulate protein function and are often altered in neurodegenerative diseases.
3. Colocalisation studies: By using multiple antibodies conjugated to different fluorescent dyes, scientists can investigate the colocalisation of different molecules, providing insights into their potential interactions or shared functions.
4. Disease markers: Immunofluorescence staining can be used to visualise pathological hallmarks of neurodegenerative diseases, such as protein aggregates in ALS.
5. Immunofluorescence Staining: Advancing Our Understanding of Neurodegenerative Diseases

In the context of neurodegenerative diseases like ALS, immunofluorescence staining has been instrumental in:

1. Characterising disease pathology: Immunofluorescence staining has been used to visualise pathological hallmarks of ALS, such as aggregates of misfolded proteins, providing insights into disease mechanisms.
2. Investigating disease mechanisms: By visualising specific molecules or processes, immunofluorescence staining has helped dissect the molecular and cellular mechanisms underlying ALS.
3. Evaluating therapeutic efficacy: Immunofluorescence staining can be used to assess the effects of potential therapeutic interventions, such as changes in the levels or localisation of a specific protein.

## Conclusion

Immunofluorescence staining has proven to be an indispensable tool in neuroscience research, enabling scientists to visualise specific molecules within cells or tissues and providing invaluable insights into the molecular and cellular processes underlying neuronal function and dysfunction. Through its various applications, this powerful technique continues to advance our understanding of the pathogenesis of neurodegenerative diseases like ALS and inform the development of novel therapeutic strategies.

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*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post explores the principles and applications of immunofluorescence staining in neuroscience research, highlighting its critical role in investigating the molecular and cellular processes underlying neuronal function and dysfunction, and advancing our understanding of neurodegenerative diseases like ALS.

Immunofluorescence Staining in Neuroscience Research

🕒 *Approximate reading time: 3 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Today in the lab was primarily a day of analysis, dedicated to the computer, scrutinising the electrical activity of neurons that I had previously recorded during my electrophysiology experiments (you can refer to my previous post here). Today's agenda involved a considerable amount of coding via Matlab software, to compute various parameters like amplitude, frequency, and charge of the neuronal electrical activity. Performing these calculations necessitates a range of intricate mathematical functions, but the procedure is semi-automated thanks to computer code and software specifically designed for this type of analysis. The final product? Intricate graphs visually demonstrating the different components of the data analysis, as depicted below.

![{595x714} Detection of mEPSCs in a recording from a neuron. (A) Sum of two exponentials template (red) and mean event overlay (green). (B) Deconvoluted wave of the recording (blue), detection threshold (green), and detected events (red dots). (C) All-point histogram of the deconvoluted trace (blue bars) fitted with a Gaussian function (red curve). The detection threshold (green) was set to 4x the standard deviation of the Gaussian. (D) Continuous trace of detected mEPSCs (red). (E) Superimposed detected mEPSCs (grey curves) and the median mEPSC (black curve) during a continuous recording, aligned horizontally to the peak of the deconvolution function. (F) Merged events from entire recording wave with the mean ensemble (blue) and fit (red).](/media/ephys.png "Data analysis")

## An Insight into Today's Data Analysis

Miniature excitatory postsynaptic currents (mEPSCs; essentially the basal electrical activity of neurons) were examined using a deconvolution-based method specifically developed for detecting spontaneous mEPSCs. This approach entails fitting events with a sum of two exponential functions (one each for rise and decay). This template is generated based on the time course of spontaneous synaptic events in the recorded waves, which are then deconvolved from the template using Fourier transformation algorithms. The deconvoluted wave is fitted with a Gaussian function, representing the noise distribution, and the detection threshold is set to four times the standard deviation of this Gaussian. The deconvoluted trace is subsequently scanned for local maxima, which correspond to the detected mEPSCs and their onset times. Subsequently, the median amplitude, frequency, charge, rise time and decay time were calculated for each individual neuron.

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*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

The blog post delves into the analytical side of neuroscience, exploring the process of examining neuronal electrical activity using Matlab software and a host of complex mathematical functions, complete with data visualisations.

A Day in the Lab: Analysing the Electrical Activity of Neurons using Computer Code

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Mitochondria, often known as the powerhouses of the cell, are at the centre of cellular health and disease. In recent years, a mounting body of evidence implicates mitochondrial dysfunction in the progression of neurodegenerative diseases.

## The Role of Mitochondria in Cells

Mitochondria are responsible for producing the majority of the energy cells need to function, in the form of a molecule called adenosine triphosphate (ATP). Moreover, they play a role in many other cellular processes, including calcium homeostasis, reactive oxygen species production, and programmed cell death or apoptosis.

## Mitochondrial Dysfunction and Neurodegenerative Diseases

Mitochondrial dysfunction has been linked to a range of neurodegenerative diseases. Below are a few examples:

1. Alzheimer's Disease: Abnormal mitochondrial dynamics and function have been observed in both cellular and animal models of Alzheimer's. Moreover, certain genetic mutations associated with familial Alzheimer's disease have been shown to impact mitochondrial function.

2. Parkinson's Disease: Genetic studies have identified mutations in mitochondrial genes in familial forms of Parkinson's disease. Moreover, a hallmark of the disease, the loss of dopaminergic neurons, has been associated with mitochondrial dysfunction.

3. Amyotrophic Lateral Sclerosis (ALS): In ALS, there's evidence of mitochondrial damage and dysfunction in the affected motor neurons.

## The Mechanisms of Mitochondrial Dysfunction in Neurodegeneration

Research is still ongoing to uncover how mitochondrial dysfunction contributes to neurodegeneration. Several mechanisms are under investigation:

1. Energy Depletion: If mitochondria fail to produce sufficient ATP, neurons may not function properly and could eventually die.

2. Oxidative Stress: Mitochondria are a major source of reactive oxygen species, which can damage cells if not properly managed.

3. Calcium Dysregulation: Abnormal mitochondrial handling of calcium can disrupt neuronal communication and trigger cell death.

## The Promise of Targeting Mitochondria in Therapies

Given the role of mitochondrial dysfunction in neurodegenerative diseases, targeting mitochondria to improve their function or protect them from damage holds therapeutic promise. Various strategies are under development, including enhancing mitochondrial biogenesis, improving mitochondrial quality control mechanisms, and protecting against oxidative damage.

## Conclusion

While there's much still to learn, the connection between mitochondrial dysfunction and neurodegenerative diseases is clear. By furthering our understanding of this relationship, we can uncover new strategies to tackle these devastating conditions that affect millions around the world.

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*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post explores the intriguing link between mitochondrial dysfunction and neurodegenerative diseases. We delve into the roles of mitochondria in cells, examine the evidence connecting mitochondrial dysfunction to neurodegeneration, and consider the promising therapeutic strategies targeting mitochondria.

The Role of Mitochondria in Neurodegenerative Diseases

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

The blood-brain barrier (BBB) is a unique feature of our biology. This protective boundary has a crucial role in maintaining the stable environment needed for the brain to function correctly, and a malfunctioning BBB can have significant implications for brain health.

## What is the Blood-Brain Barrier?

The BBB is a semipermeable border that separates the circulating blood from the brain and extracellular fluid in the central nervous system (CNS). Composed primarily of endothelial cells lining the blood vessels, the BBB also involves other components such as astrocytes and pericytes, forming what's known as the neurovascular unit.

## The Function of the Blood-Brain Barrier

The BBB has several crucial functions:

1. Protection: It protects the brain from harmful substances in the blood, such as toxins, pathogens, and some drugs, by preventing their passage into the brain.

2. Homeostasis: It maintains the chemical composition of the brain's extracellular fluid, providing a stable environment for neuronal function.

3. Nutrient Transport: It selectively transports essential nutrients and molecules from the blood into the brain.

4. Neuronal Activity Regulation: It helps regulate neuronal activity by controlling the supply of hormones and nutrients and removing waste products.

## The Blood-Brain Barrier and Disease

Dysfunction or breakdown of the BBB is associated with several neurological conditions:

1. Multiple Sclerosis: The disruption of the BBB allows immune cells to enter the CNS, causing inflammation and damage.

2. Alzheimer's Disease: Some research suggests that BBB breakdown may contribute to the buildup of beta-amyloid plaques characteristic of Alzheimer's disease.

3. Stroke: In stroke, the BBB is often damaged, leading to brain oedema and worsening the injury.

4. Brain Tumours: BBB can restrict the delivery of therapeutic agents to brain tumours, posing a significant challenge in their treatment.

## Future Directions and Conclusion

Understanding the BBB's functions and its role in disease can help develop strategies to protect it or selectively bypass it for drug delivery. Emerging techniques such as nanoparticle-based drug delivery systems and focused ultrasound are showing promise in this regard.

As we continue to unravel the complexities of the BBB, we grow closer to developing effective treatments for devastating neurological diseases that currently have limited therapeutic options.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post delves into the fascinating world of the blood-brain barrier. We explore its function, its importance in maintaining brain health, its involvement in various neurological conditions, and the potential future directions of research in this field.

The Blood-Brain Barrier: Understanding Its Function and Importance

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Today's adventure in the lab revolved around an electrophysiology experiment, with a particular emphasis on the patch-clamp technique. The underlying concept involves utilising an electrode to record the electrical activity of a solitary neuron grown in vitro. Admittedly, the procedure can initially seem somewhat challenging, but with practice, it becomes significantly more manageable. Over the past two months, I have successfully recorded activity from roughly 100 individual neurons, each requiring approximately 20 minutes of dedicated attention.

Presented below are images that depict what the neurons appeared like under the microscope. Further details on the experiment and the methods I employed are detailed subsequently.

![{596x466} These are some of the neurons from which I recorded electrical activity today. Image (A) illustrates a neuron under a DIC microscope. Images B-D display neurons labelled with a fluorescent reporter.](/media/ephys2.png "Neurons")

To comprehend how neuronal activity is recorded and how to operate the recording equipment, one requires a solid understanding of physics and electronics. Electrophysiology demands it. Below, I've included an image of the various recording equipment utilised in today's experiment. The major components are labelled for the convenience of the curious among you. The setup principally involves a DIC (differential interference contrast) microscope connected to a range of electronic devices.

![{1270x638} The array of equipment employed for my experiment.](/media/ephys0.png "Electrophysiology equipment")

## Delving into the Methodology of Today's Experiment

The recordings were performed employing a computer-controlled patch clamp amplifier, which was operated with specialised computer software. Whole-cell voltage clamp recordings were performed at a holding potential of -65 mV at room temperature. The whole-cell configuration was achieved by applying a gentle suction pulse orally while delivering a brief 1 V electrical pulse simultaneously. Compensation was applied for the fast and slow capacity transients. A seal test (a 10 mV test pulse) was administered through the recording electrode to measure the access resistance at the commencement of every 10-second recording.

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

Join me on a fascinating journey in the lab as I employ the sophisticated patch-clamp technique in an electrophysiology experiment to record neuronal activity.

A Day in the Lab: Unravelling Neuronal Activity using Electrophysiology

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Oxidative stress, an imbalance between the production of free radicals and the ability of the body to counteract their harmful effects, is becoming increasingly recognised as a crucial player in the health of neurons and the pathogenesis of neurodegenerative diseases.

## Understanding Oxidative Stress

Oxidative stress arises when there's an imbalance between the production of free radicals, such as reactive oxygen species (ROS), and the body's ability to detoxify these reactive intermediates or repair the resulting damage. These free radicals are highly reactive due to unpaired electrons and can cause oxidative damage to DNA, proteins, and lipids, potentially leading to cell death.

## Oxidative Stress and Neuronal Health

Neurons are particularly susceptible to oxidative stress due to their high metabolic rate, their reliance on oxidative phosphorylation for energy production, and their relatively low antioxidant capacity. Oxidative damage in neurons can lead to functional impairment and ultimately neuronal death.

## Oxidative Stress and Neurodegenerative Diseases

Oxidative stress has been linked to several neurodegenerative diseases:

1. Alzheimer's Disease: Oxidative damage to proteins, lipids, and DNA has been observed in the brains of patients with Alzheimer's disease.

2. Parkinson's Disease: There is evidence of oxidative damage in the substantia nigra, a region of the brain affected in Parkinson's disease.

3. Amyotrophic Lateral Sclerosis (ALS): Oxidative damage is also implicated in ALS, with increased levels of oxidative markers found in patients.

## Targeting Oxidative Stress in Neurodegenerative Diseases

Given the role of oxidative stress in neurodegeneration, strategies to combat oxidative stress are being explored as potential treatments for these diseases. This could involve antioxidants that directly neutralise free radicals or drugs that enhance the body's own antioxidant defences.

## Conclusion

While we are still in the early stages of understanding the complex role of oxidative stress in neuronal health and disease, it's clear that this cellular phenomenon plays a critical role in neurodegeneration. Future research aimed at further elucidating the molecular mechanisms of oxidative stress and developing ways to counteract it may hold the key to novel therapies for neurodegenerative diseases.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

In this blog post, we delve into the impact of oxidative stress on neuronal health. We cover what oxidative stress is, how it affects neurons, its role in neurodegenerative diseases, and potential strategies for targeting oxidative stress in these conditions.

The Impact of Oxidative Stress on Neuronal Health

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Genotyping, the process of determining an organism's genetic variants, has been pivotal in unveiling the genetic basis of neurological disorders like amyotrophic lateral sclerosis (ALS). This blog post will delve into the importance of genotyping in the field of neuroscience and how it informs our understanding of neurodegenerative diseases.

## Genotyping: What Does It Involve?

Genotyping involves the use of molecular biology techniques to determine an individual's specific sequence of genes or genetic markers. Common genotyping techniques include Polymerase Chain Reaction (PCR), DNA sequencing, and microarray-based methods. The specific technique chosen largely depends on the type of genetic variant (such as single-nucleotide polymorphisms or copy number variations) that researchers are interested in.

## Implications of Genotyping in Neuroscience

Genotyping has several applications in neuroscience research:

1. Understanding Genetic Basis of Diseases: Many neurological disorders, such as ALS, Parkinson's, and Alzheimer's, have a genetic component. Genotyping can help identify the specific genes and genetic variants associated with these diseases.

2. Predicting Disease Risk: Some genetic variants can increase an individual's risk of developing certain neurological disorders. By identifying these risk variants through genotyping, we can predict disease risk more accurately.

3. Personalised Medicine: Understanding an individual's genetic makeup can help inform treatment decisions, such as predicting their response to specific drugs, a concept known as personalised or precision medicine.

## Genotyping in Neurodegenerative Diseases

In the context of neurodegenerative diseases, genotyping has been instrumental in:

1. Identifying Disease-Causing Mutations: Several neurodegenerative diseases are associated with specific genetic mutations. For example, mutations in the SOD1 gene were the first to be linked to ALS.

2. Disease Modelling: Genotyping is essential in creating animal models of neurodegenerative diseases. Researchers can engineer animals to carry disease-causing mutations and then study the disease progression and potential treatments in these models.

3. Screening and Diagnostic Purposes: Genotyping can be used to identify carriers of disease-causing mutations, particularly in families with a history of a specific neurodegenerative disease. This can help in early diagnosis and intervention.

## Conclusion

Genotyping serves as a critical tool in neuroscience, enhancing our understanding of the genetic underpinnings of neurological disorders. It is a powerful technique that holds promise for the development of novel diagnostic tools and treatments tailored to an individual's genetic profile, pushing us towards a future of personalised medicine in neurology.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post explores the role of genotyping in neuroscience, highlighting its importance in understanding the genetic basis of neurological disorders, predicting disease risk, and informing personalised treatment strategies.

Genotyping: Unravelling the genetic basis of neurological disorders

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Polymerase chain reaction (PCR) is an indispensable technique in neuroscience, allowing researchers to amplify and analyse gene expression in neural tissues or cultured cells. Understanding the application of PCR in neuroscience opens doors to studying genetic influences on the nervous system and how they contribute to neurological diseases like amyotrophic lateral sclerosis (ALS).

## The Basics of PCR

PCR is a technique used to amplify specific DNA sequences. It involves three primary steps:

1. Denaturation: The DNA sample is heated to separate the two DNA strands.

2. Annealing: The temperature is lowered to allow primers, short DNA sequences, to bind to their complementary sequences on the DNA strands.

3. Extension: A heat-tolerant DNA polymerase enzyme extends the primers, synthesising new strands of DNA.

These steps are repeated for multiple cycles, each one doubling the amount of DNA, leading to exponential amplification of the target sequence.

## Applications of PCR in Neuroscience

1. Studying Gene Expression: With reverse transcription PCR (RT-PCR), scientists can quantify mRNA levels, allowing for the study of gene expression in different neuronal types or under different conditions, such as disease states or drug treatments.

2. Genotyping Animals: PCR can be used to identify transgenic animals or those carrying specific gene mutations, both of which are commonly used in neuroscience research.

3. Identifying Pathogen Infection: PCR can detect the presence of pathogen DNA or RNA in brain tissue, helping identify viral or bacterial infections that could affect neurological function.

## PCR in Neurodegenerative Disease Research

In studying neurodegenerative diseases like ALS, PCR offers several advantages:

1. Analysing Disease-Associated Genes: PCR allows for the study of mutations in disease-associated genes, such as the SOD1 gene in ALS.

2. Studying Gene Expression Changes: Using RT-PCR, researchers can explore how disease progression affects the expression of genes involved in neuroinflammation, neuronal survival, or other relevant processes.

3. Evaluating Therapeutic Efficacy: Following gene therapy or other genetic interventions, PCR can help assess whether the treatment has effectively altered gene expression.

## Conclusion

PCR is a powerful tool in neuroscience, enabling the amplification and analysis of specific genes. It allows researchers to delve into the genetic underpinnings of neuronal function and dysfunction, which is essential for understanding neurological diseases and developing effective treatments. Its versatility and applicability make PCR a cornerstone technique in neuroscience research.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post provides an overview of the role of PCR in neuroscience, explaining how it allows for the amplification and analysis of gene expression in studying neurological function and disease.

PCR in Neuroscience: Amplifying and Analysing Gene Expression

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

The marvels of neuronal communication have long captivated neuroscientists. Thanks to the development of calcium imaging techniques, we're now able to witness this cellular dialogue in real time, enhancing our comprehension of brain activity.

## Understanding Calcium's Role in Neurons

Calcium ions (Ca2+) play a crucial role in neurons. Upon neuronal activation, there is an influx of Ca2+ into the cell. This calcium influx triggers a cascade of events, including the release of neurotransmitters – the chemical messengers that allow neurons to communicate.

## What is Calcium Imaging?

Calcium imaging is a technique that enables us to visualise the activity of neurons by detecting changes in intracellular Ca2+ levels. This is typically achieved using fluorescent dyes or proteins that bind to Ca2+ and emit light when they do so. By monitoring these changes in fluorescence, we can infer when and where neuronal activity is taking place.

## Applications of Calcium Imaging in Neuroscience

Calcium imaging offers a powerful way to study the brain and its functions:

1. Understanding Neuronal Networks: By visualising the activity of individual neurons, we can gain insights into how neurons work together in networks.

2. Unveiling the Mechanisms of Learning and Memory: Changes in the strength of connections between neurons, which is influenced by Ca2+ signalling, underlie learning and memory. Calcium imaging can help reveal these processes.

3. Studying Neurological and Neurodegenerative Diseases: Abnormal Ca2+ signalling is implicated in several neurological and neurodegenerative disorders. Using calcium imaging, we can investigate these abnormalities in models of diseases such as Alzheimer's and Parkinson's.

## The Future of Calcium Imaging

Recent advances are pushing the boundaries of calcium imaging. For instance, two-photon microscopy allows for calcium imaging deep within brain tissue, and the development of genetically encoded calcium indicators is enabling long-term imaging in specific types of neurons. These innovations are opening new avenues for exploring the intricate symphony of neuronal communication.

## Conclusion

From understanding basic neuronal function to unravelling complex cognitive processes and studying neurological diseases, calcium imaging serves as an invaluable tool in neuroscience research. As technology advances, the precision and applicability of calcium imaging will only increase, illuminating the mysteries of the brain like never before.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post sheds light on calcium imaging, a powerful technique for capturing the dynamic dialogue between neurons. We explore the fundamental role of calcium in neurons, delve into the process of calcium imaging, and consider its significant applications and future in neuroscience research.

Calcium imaging: Capturing neuronal communication

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Neuronal culture, the practice of growing neurons outside the body, is an invaluable method in neuroscience research. By observing neurons in isolation, scientists can study their individual behaviours, physiological properties, and responses to various stimuli more effectively than in whole animals or human subjects.

## Steps in Preparing Neuronal Cultures

1. Tissue Dissection and Dissociation: Brain or spinal cord tissue, often from embryonic or neonatal rodents, is dissected and dissociated into individual cells. The process involves a combination of mechanical trituration and enzymatic digestion.

2. Plating and Initial Culture: Cells are plated onto a substrate coated with materials like poly-D-lysine or laminin that promote adhesion. The cells are cultured in a suitable medium that supports neuronal survival and growth.

3. Maintenance: Neurons require careful maintenance, including regular medium changes and monitoring of their health and growth.

## Challenges in Culturing Neurons

1. Neurons are post-mitotic and sensitive cells, which poses specific challenges for their culture:

2. Survival and Growth: Neurons have high metabolic demands and require specific growth factors and nutrients, often provided by glial cells in vivo. Co-culture with glial cells or supplementation with specific factors can support neuronal survival and growth in culture.

3. Neuronal Polarity: In vivo, neurons develop a highly polarised morphology with distinct axons and dendrites. Reproducing this polarity in culture can be challenging.

4. Connectivity: In vivo, neurons form complex networks of synaptic connections. In culture, neurons can form synapses, but reproducing the complex connectivity of the brain is not yet feasible.

## Applications of Neuronal Culture in Neuroscience

Despite these challenges, neuronal cultures have proved instrumental in studying various aspects of neuroscience:

1. Neuronal Development: By observing neurons in culture, researchers can study processes like axon guidance, dendrite branching, and synapse formation in detail.

2. Neuronal Physiology: Neuronal cultures allow for detailed studies of neuronal electrophysiology, including action potential firing and synaptic transmission.

3. Disease Modelling: Neurons derived from patients or genetically engineered models can be used to study disease mechanisms and test potential therapeutics.

## Neuronal Culture in the Study of Neurodegenerative Diseases

In the context of neurodegenerative diseases like ALS, neuronal cultures offer several advantages:

1. Disease Mechanisms: Culturing neurons expressing disease-associated mutations can help researchers understand how these mutations disrupt neuronal function.

2. Drug Testing: Potential therapeutics can be tested on cultured neurons to assess their effects on neuronal health and function.

3. Patient-Specific Models: With the advent of induced pluripotent stem cells (iPSCs), neurons can be derived from patients, allowing for the study of disease mechanisms in a patient-specific context.

## Conclusion

Neuronal culture, while not without its challenges, is a powerful tool in neuroscience. It allows researchers to observe neurons in isolation, providing unique insights into neuronal development, function, and disease mechanisms. The continued refinement of neuronal culture techniques will undoubtedly contribute to our understanding of the nervous system and the development of therapeutics for neurodegenerative diseases like ALS.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post explores the methods and challenges of neuronal culture, demonstrating its crucial role in studying neuronal development, function, and disease mechanisms.

Neuronal Culture: Growing and Maintaining Healthy Neurons

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Viral vectors have revolutionised neuroscience research. They serve as tools to introduce foreign genetic material into cells, thereby enabling a plethora of experimental manipulations, from labelling specific neurons to modifying their function.

## What are Viral Vectors?

A viral vector is a virus that has been engineered to carry foreign genetic material into a host organism’s cells. These vectors capitalise on the natural ability of viruses to infect cells and hijack their machinery to replicate. However, in the context of research, viral vectors are modified to remove their disease-causing ability and to incorporate the genetic material of interest.

## Commonly Used Viral Vectors in Neuroscience

Different viral vectors are used in neuroscience research, each with its unique advantages:

Adenoviruses: Known for their high transduction efficiency and ability to infect both dividing and non-dividing cells, they're often used for short-term studies due to their transient gene expression.

Lentiviruses: These can infect both dividing and non-dividing cells and provide long-term gene expression, making them ideal for studies requiring lasting changes.

Adeno-associated viruses (AAVs): They are one of the most frequently used vectors in neuroscience due to their low immune response, long-term gene expression, and broad range of serotypes allowing specific targeting of different cell types.

## Applications of Viral Vectors in Neuroscience

The use of viral vectors in neuroscience is vast and varied:

1. Gene Therapy: Viral vectors can deliver genes to replace or supplement defective ones in neurodegenerative disorders, offering promise for therapeutic applications.

2. Neuronal Labelling: Viral vectors can introduce fluorescent proteins into neurons, allowing scientists to visualise and trace neural circuits.

3. Optogenetics: By delivering genes that encode for light-sensitive proteins, scientists can use light to control neuronal activity, a field known as optogenetics.

4. Chemogenetics: Viral vectors are used to introduce receptors that can be activated by specific drugs, allowing precise control over neuronal activity.

## The Future of Viral Vectors in Neuroscience

As we refine our ability to manipulate viral vectors and enhance their specificity, we open up new possibilities for understanding and treating brain disorders. From targeted therapies for neurodegenerative diseases to the revelation of complex neuronal circuits, the future of neuroscience is inextricably tied to these microscopic tools.

## Conclusion

Viral vectors have dramatically expanded our capacity to explore the intricate workings of the brain. They exemplify the symbiotic relationship between neuroscience and virology, where our understanding of viruses has been harnessed to deepen our knowledge of the brain and its myriad disorders.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post delves into the exciting world of viral vectors and their transformative role in neuroscience research. We explore their definition, types, applications, and future prospects, shedding light on how these microscopic tools are shaping our understanding of the brain.

Viral Vectors in Neuroscience Research

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Epigenetics, the study of changes in gene expression without alterations in the DNA sequence, is shedding new light on the complex mechanisms underlying neurodegenerative diseases.

## Understanding Epigenetics

Epigenetics involves changes to gene expression that do not involve alterations to the underlying DNA sequence. These changes include DNA methylation, histone modification, and non-coding RNA molecules, which can all alter the structure of the chromatin and subsequently regulate gene expression.

## Epigenetics and Neurodegeneration

A growing body of research suggests that epigenetic alterations may play a crucial role in neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis.

1. Alzheimer's Disease: Research has shown that epigenetic changes, particularly DNA methylation and histone modifications, may be involved in the pathogenesis of Alzheimer's disease.

2. Parkinson's Disease: Aberrant DNA methylation and changes in histone modifications have been reported in patients with Parkinson's disease.

3. Amyotrophic Lateral Sclerosis: Altered expression of non-coding RNAs has been implicated in the pathogenesis of ALS.

## Epigenetic Therapies in Neurodegeneration

Given the potential role of epigenetics in neurodegenerative diseases, there's growing interest in developing epigenetic therapies. These therapies aim to modify the epigenetic state of specific genes or genomic regions, with the hope of halting or even reversing disease progression.

## Challenges and Future Directions

However, the development of epigenetic therapies faces several challenges. The brain is a highly complex organ with many different cell types, each with its own epigenetic landscape. Targeting the epigenetic changes in the correct cell type without affecting others is a significant challenge.

## Conclusion

While we are still in the early stages of understanding the complex relationship between epigenetics and neurodegeneration, it's clear that this is a promising area of research. Further exploration of the epigenetic landscape in neurodegenerative diseases could pave the way for novel therapeutic strategies.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post explores the complex relationship between epigenetics and neurodegeneration. We cover what epigenetics is, how it might contribute to neurodegenerative diseases, and the potential for epigenetic therapies in these conditions.

Epigenetics and Neurodegeneration: A Complex Relationship

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

In the realm of neuroscience, patch-clamp electrophysiology has emerged as an indispensable tool for studying neurons and their electrical activity. This technique offers an in-depth look into the intricate processes that allow neurons to communicate, thus propelling our understanding of brain function.

## What is Patch-Clamp Electrophysiology?

Patch-clamp electrophysiology is a technique that allows the study of single or multiple ion channels in cells. It involves the use of a glass micropipette, which is attached to the membrane of a cell, forming a 'patch'. Once the patch is formed, it can be used to measure the electrical activity within the cell or across its membrane.

## Types of Patch-Clamp

There are different configurations of patch-clamp electrophysiology, each serving unique purposes:

1. Cell-attached: The pipette is attached to the cell membrane, allowing the study of the activity of individual ion channels within the patch.

2. Whole-cell: The pipette breaks the cell membrane, providing a direct electrical link to the interior of the cell and enabling the measurement of overall electrical activity.

3. Inside-out and outside-out: In these configurations, the pipette is retracted after forming a cell-attached patch, pulling a patch of membrane with it. This allows the exposure of intracellular or extracellular face of the membrane to different solutions, aiding the study of ion channel regulation.

## Applications of Patch-Clamp Electrophysiology

Patch-clamp electrophysiology has several applications in neuroscience:

1. Understanding Neuronal Function: It allows researchers to measure the electrical activity of neurons and understand the role of different ion channels in this activity.

2. Drug Development: By studying the effect of potential therapeutic drugs on neuronal activity, researchers can evaluate their efficacy and mechanism of action.

3. Studying Neurological Disorders: Certain neurological disorders are associated with abnormal ion channel function. Patch-clamp techniques can be used to understand these abnormalities.

## Conclusion

Patch-clamp electrophysiology has revolutionised our ability to study the electrical activity of neurons at an unprecedented level of detail. As we continue to refine and develop this technique, it will undoubtedly provide further insights into neuronal function and dysfunction, with profound implications for our understanding of the brain and the treatment of neurological disorders.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post delves into the world of patch-clamp electrophysiology, a technique that offers an unparalleled view into the electrical activity of neurons. We explore the different configurations of the technique, its applications in neuroscience, and its transformative role in the study of brain function.

Patch-clamp electrophysiology: Measuring neuronal activity

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Neuroimaging techniques have opened the door to the brain, allowing us to visualise its structure and function in unprecedented detail. These technologies have transformed neuroscience, enabling us to explore the brain in health and disease, and providing insights that extend from the level of individual neurons to complex neural circuits.

## Basics of Neuroimaging

Neuroimaging refers to the use of various techniques to directly or indirectly image the structure, function, or pharmacology of the nervous system. It is a relatively new discipline within medicine and neuroscience, and combines various technologies to visualise the nervous system in real time.

## Structural and Functional Neuroimaging

Structural neuroimaging provides information about the anatomical structure of the brain, allowing us to visualise different brain areas and detect any structural changes or abnormalities. Techniques include computed tomography (CT), magnetic resonance imaging (MRI), and diffusion tensor imaging (DTI).

Functional neuroimaging, on the other hand, provides information about brain activity, letting us see which brain areas are active during different tasks or mental states. These techniques include functional magnetic resonance imaging (fMRI), positron emission tomography (PET), and electroencephalography (EEG).

## Applications of Neuroimaging in Neuroscience

1. Understanding Brain Function: Neuroimaging allows us to identify which brain areas are involved in different cognitive processes, such as memory, attention, or emotion.

2. Diagnosing Brain Disorders: Neuroimaging can be used to identify structural or functional abnormalities in the brain associated with disorders such as stroke, tumours, or neurodegenerative diseases.

3. Tracking Disease Progression: In diseases like Alzheimer's or Parkinson's, neuroimaging can help track the progression of the disease and the effects of therapeutic interventions.

## Neuroimaging in Neurodegenerative Diseases

Neuroimaging has become an essential tool in studying neurodegenerative diseases:

1. Detecting Early Changes: For example, structural MRI can detect brain shrinkage in early stages of Alzheimer's disease, before symptoms appear.

2. Understanding Disease Mechanisms: Functional imaging can help reveal how neurodegenerative diseases alter brain activity.

3. Evaluating Treatments: Neuroimaging can be used to track how experimental treatments affect the brain, providing important information for drug development.

## Conclusion

Neuroimaging has revolutionised our ability to visualise the living brain, deepening our understanding of brain function and disease. As these technologies continue to advance, they will undoubtedly continue to illuminate the complexities of the brain in increasingly sophisticated ways.

-----

*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post explores the remarkable capabilities of neuroimaging techniques in visualising the living brain, examining both structural and functional imaging approaches, their applications in neuroscience, and their indispensable role in the study of neurodegenerative diseases.

Neuroimaging Techniques: Visualising the Living Brain

🕒 *Approximate reading time: 2 minutes*

✍️ *Author: Dr Eleni Christoforidou*

I am thrilled to share that the University of Glasgow's alumni magazine, 'Avenue', reached out to me recently for an interview. They wanted to delve into my journey as both a student resident and a Living Support Assistant in the university halls.

As a final year student, I have spent my entire academic tenure living within the university's residential facilities. I have been lucky enough to witness the university's thriving student life from a unique perspective. However, my relationship with the halls goes beyond just living there. For the past three years, I have been serving as a Living Support Assistant. This role has allowed me to further connect with our diverse student community and contribute to their experience within the university halls.

The interview with 'Avenue' was a fantastic opportunity to reflect on my experiences and share some insights about university life that I've gathered over the years. I spoke about the camaraderie and friendships formed within the halls, the challenges that students often face while adjusting to university life, and how we, as Living Support Assistants, work to ensure that the student experience is as comfortable and enriching as possible.

One of my most cherished aspects of this role has been seeing students grow and evolve during their time at the university. Being able to provide guidance and support during their transitional phase has been incredibly rewarding. Not only has it given me a sense of purpose, but it's also enriched my own student experience by allowing me to understand different perspectives and cultures.

The interview also brought back fond memories of the various events we organised in the halls. From fresher's welcome parties to cultural nights and study support sessions, these events have been instrumental in fostering a sense of community and belonging among the students.

Being recognised by 'Avenue' has been a delightful surprise and an honour, to say the least. It's a testament to the amazing work we are doing within the university halls and a reflection of the supportive and vibrant community that the University of Glasgow nurtures.

You can read a snippet of the mini interview below.

![{1170x846}](/media/annotation-2019-09-14-195035.png "Alumni magazine interview 2018")


In this blog post, I share my unique journey as a student and Living Support Assistant in the University of Glasgow's halls of residence, and my excitement about being interviewed for the university's alumni magazine, 'Avenue'.

My interview for Glasgow University's Alumni Magazine

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Today, I've been engrossed in studying H&E stained brain slices, attempting to unearth histopathological modifications in the aftermath of a stroke. H&E-stained brain tissue was scrutinised under a light microscope. The somas of healthy, unaffected neurons exhibited a round shape, with a distinctly visible nucleus. Contrarily, ischaemic neurons displayed a dark and shrunken appearance, coupled with pyknotic nuclei. Moreover, eosin staining appeared more muted and there was noticeable vacuolation of the neuropil. A significant infiltration of neutrophils was observed, and oedema was glaringly apparent. Below, I've included some of the images I captured using the microscope.

![Microscopy](/media/Light-Microscopy.mp4 "Microscopy")

_Timelapse: Observing the stained brain slices under a light microscope._

![{1190x440} Light microscopic images (x40 magnification) of H&E-stained brain tissue post-stroke. The left image presents unaffected tissue from the ipsilateral hemisphere. The right image exhibits the infarct.](/media/h-e-staining.png "H&E staining")

![{840x479} T2-weighted MRI images, 7 days post-stroke, alongside diagrams of corresponding coronal slices. The infarct area, observed via light microscopic examination of H&E-stained sections, was manually drawn onto the diagrams and is showcased in red.](/media/annotation-2020-01-11-153925.png "MRI images")

## Unpacking the Methodology of Today's Experiment

The infarct area, as observed on H&E-stained sections under the light microscope, was manually transcribed onto diagrams of corresponding coronal slices from a brain atlas. These were subsequently scanned into a computer and scrutinised using image analysis software. The area of the drawn infarct was manually delineated, mirroring the method applied for T2-weighted MRI images, but without oedema correction. The infarct area from each diagram was compared with the oedema-corrected and uncorrected infarct areas from the corresponding slices from T2-weighted MRI images of the same brain, to evaluate the precision of infarct measurement from MRI images. Oedema-corrected infarct areas from T2-weighted images were calculated using formulas from previously published research. Lesion areas deduced from diagrams were significantly smaller than those calculated from T2-weighted images prior to oedema correction.

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*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

Join me as I delve into a day in the lab, analysing H&E stained brain slices to unravel the histopathological transformations following a stroke.

A Day in the Lab: Decoding the Histopathological Alterations in Brain Slices Post-Stroke

🕒 *Approximate reading time: 4 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Optogenetics is a revolutionary technique in neuroscience that combines genetics and optics to control and observe specific events in the cells of living tissue. It is particularly powerful in the study of neurons, where it enables researchers to control the activity of individual neurons and observe the consequences on the nervous system in real time.

## The Basics of Optogenetics

Optogenetics involves the introduction of light-sensitive proteins, often ion channels, into specific cells. These proteins, derived from microorganisms, respond to light of specific wavelengths by opening or closing, which can trigger or inhibit neuronal firing.

This technique requires the use of genetic methods to introduce the gene coding for the light-sensitive protein into the neurons of interest. This is often achieved via viral vectors or transgenic animals.

Once these light-sensitive proteins are expressed in the neurons, a light source (like a laser) is used to control the activity of these neurons. This manipulation can be done with incredible precision, both in terms of the specific neurons targeted and the timing of their activation or inhibition.

## Applications of Optogenetics in Neuroscience

1. Unravelling Neural Circuits: Optogenetics allows researchers to activate or inhibit specific neurons within a circuit and observe the consequences, thus helping unravel the complex workings of neural circuits.

2. Decoding Neuronal Coding: By manipulating neuronal activity and observing the consequences on an animal's behaviour or perception, researchers can begin to decode how neurons encode information about the world.

3. Developing New Therapies: Optogenetic control of neurons could potentially be used as a therapy in conditions like blindness, Parkinson's disease, or epilepsy.

## Optogenetics in Neurodegenerative Diseases

While most applications of optogenetics are in basic neuroscience, there are emerging applications in the study of neurodegenerative diseases:

1. Understanding Disease Mechanisms: By enabling precise control over neuronal activity, optogenetics can be used to study how changes in neuronal function contribute to neurodegenerative diseases.

2. Testing Therapeutic Approaches: Optogenetics could potentially be used to test the effects of increasing or decreasing activity of specific neuronal populations in animal models of neurodegenerative diseases.

## Conclusion

Optogenetics has revolutionised our ability to study neurons and neural circuits with an unprecedented level of precision. Its potential applications, from understanding basic neuronal function to developing new therapies for neurodegenerative diseases, make it an exciting area of neuroscience research.

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*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

This blog post delves into the innovative technique of optogenetics in neuroscience, discussing how it enables precise manipulation of neuronal activity, thereby aiding in the understanding of neural circuits, decoding of neuronal coding, and the development of novel therapeutic approaches.

Optogenetics: Illuminating neuronal function

🕒 *Approximate reading time: 1-2 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Today's scientific endeavour in the laboratory centred on the task of staining brain sections utilising Hematoxylin & Eosin (H&E) in preparation for microscopy. Captured in the timelapse video below, this meticulous process spanned approximately 3.5 hours.

This experiment forms a pivotal aspect of my ongoing research project at the university. Our primary objective is to determine the therapeutic efficacy of a centrally-administered peptide, introduced 35 minutes post-induction of an experimental stroke model. This determination is achieved by analysing the final infarct volume and the growth of the ischaemic lesion through Magnetic Resonance Imaging (MRI). Given that the acquired MRI images, both Diffusion Weighted Imaging (DWI) and T2-weighted images, do not provide a quantifiable measure of cellular death but rather display varying patterns of oedematous activity, it was necessary to perform H&E staining. This supplementary procedure guarantees the validity of the imaging results post-oedema correction.

![H&E staining experiment](/media/lab.mp4 "H&E staining experiment")

_Timelapse: Conducting the H&E staining experiment._

## Elaborate Methodology of Today's Experiment

A grand total of 54 brain sections underwent staining. Commencing with paraffin-processed brain sections, they were submerged in histoclear, followed by sequential immersion in a series of graded alcohols. Subsequently, the application of Haematoxylin stain took place, trailed by a series of rinses and differentiation in acid alcohol. Post-dehydration, the sections were stained with eosin, proceeded by an additional round of dehydration, and finally immersed in histoclear once more. The stained sections were mounted utilising a distyrene, plasticiser, and xylene (DPX) mounting medium. Under a light microscope, basophilic structures stained by haematoxylin present in a blue shade, whereas eosinophilic structures stained by eosin adopt a pink hue. Thus, neuronal nuclei will be observed as blue, and the cytoplasm, alongside the extracellular space, will manifest as pink.

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*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

In this detailed account of a recent neuroscience experiment, I guide you through the meticulous process of staining brain slices using Hematoxylin & Eosin (H&E), shedding light on the intricate steps involved in studying the effects of a novel therapeutic approach to stroke treatment.

A Day in the Lab: Hematoxylin & Eosin Staining for Microscopical Analysis of the Brain

🕒 *Approximate reading time: 5 minutes*

✍️ *Author: Dr Eleni Christoforidou*

Today, the lab came alive with the hum of computers as I delved deep into the intricate world of image analysis. The task at hand was to sift through numerous MRI scans, collected during a trailblazing experiment designed to explore potential new treatments for stroke. I've attached an example of these images below to provide a clearer idea of the kind of analysis undertaken. Although a considerable amount of mathematics is involved, the whole process is semi-automated thanks to specialised software designed for this type of analysis. The process, albeit lengthy, offers an exciting journey through the maze of data. In fact, I anticipate spending several more days to complete the entire data set's analysis. Despite the task's enormity, the thrill of familiarising myself with the image analysis software has been a rewarding experience.

![{1002x867} This image exemplifies the image analysis I performed today. These are brain scans taken 30 minutes and 7 days post-stroke, where I've highlighted the areas of brain damage in red and white. For confidentiality reasons, I've concealed the three treatment types.](/media/brain-scans.png "Brain scans")

## Dive into the Details of Today's Image Analysis

*(Warning: You're about to dive into a pool of technical details. It's a fascinating read if you have a vested interest in the topic.)*

Today's approach to image analysis begins with identifying the initial infarct occurring 30 minutes post-stroke, defined as the hypo-intense area on quantitative apparent diffusion coefficient (ADC) maps. These maps are constructed from diffusion-weighted MRI images. To calculate the initial infarct volume for each brain, we multiplied this area on each slice by the slice thickness and then summed up the values from all eight slices.

Seven days after the stroke, the final infarct was defined by manually delineating the hyperintense area on T2-weighted MRI images. We then multiplied the infarct area from each slice by the slice thickness and summed these to calculate the lesion volume for each brain.

Ischaemic stroke, which leads to the blood-brain barrier's breakdown, results in vasogenic brain oedema due to water influx into the damaged tissue (Ayata and Ropper, 2002). This event causes an extension of the ischaemic brain lesion, accounting for about 30% of the observed infarct volume (Gerriets et al., 2004). To prevent the overestimation of infarct volume from MRI measurements, we corrected initial and final infarct volumes \[equation (1)] for swelling of the ipsilateral hemisphere using equation (2) as published in Swanson et al. (1990), and for compression of the contralateral hemisphere using equation (3) as proposed by Gerriets et al. (2004).

![Equations 1, 2, 3 {822x473}](/media/equations.png "Equations 1, 2, 3")

Subsequently, we calculated the infarct volume as a percentage of the ipsilateral hemisphere (% hemispheric lesion volume, % HLV) using equation (4) as outlined in Gerriets et al. (2004), to allow for comparison across varying total brain volumes.

![Equation 4 {581x138}](/media/equations2.png "Equation 4")

## Insights into the Experiment

A Diffusion Weighted Imaging (DWI) scan was obtained 30 minutes post-stroke to facilitate quantitative ADC map production and assessment of ischaemic tissue. The treatment was administered intracerebroventricularly via an osmotic pump inserted subcutaneously at the back of the neck, connected to a cannula inserted into the right lateral ventricle via a burr hole drilled in the contralateral skull. Post treatment, recovery was allowed for seven days, after which T2-weighted MRI images were obtained to evaluate the final infarct volume.

MRI plays a pivotal role in diagnosing and managing acute ischemic stroke. DWI, an MRI sequence that uses opposing magnetic field gradients to detect tissue in which water diffusion is restricted due to cytotoxic oedema (Fisher et al., 1992), is a reliable method of detecting acute ischaemia. Cytotoxic oedema, the swelling of neurons, glia, and endothelial cells, occurs due to the release of toxic factors after cerebral injury (Hemphill, Beal, and Gress, 2001). A heightened DWI signal in the brain is typically observed within a few minutes of stroke and leads to a reduction of the ADC value of the tissue, which quantifies the degree of water restriction (Merino and Warach, 2010).

T2-weighted imaging, another type of MRI, is employed to gauge the ischaemic lesion beyond the acute stages. T2-weighted images highlight areas of vasogenic oedema, which signify the infarct's volume. Vasogenic oedema refers to the influx of solutes and fluids into the brain due to the breakdown of the blood-brain barrier (Hemphill, Beal, and Gress, 2001).

## References

- Ayata, C. and Ropper, A.D. (2002). Ischaemic brain oedema. Journal of Clinical Neuroscience, 9, pp.113–124.
- Fisher, M., Sotak, C.H., Minematsu, K., Li, L. (1992). New magnetic resonance techniques for evaluating cerebrovascular disease. Annals of Neurology, 32, pp.115-122.
- Gerriets, T., Stolz, E., Walberer, M., Müller, C., Kluge, A., Bachmann, A., Fisher, M., Kaps, M., Bachmann, G. (2004). Noninvasive Quantification of Brain Edema and the Space-Occupying Effect in Rat Stroke Models Using Magnetic Resonance Imaging. Stroke, 35(2), pp.566-571.
- Hemphill, J.C., Beal, M.F., Gress, D.R. (2001). Critical care in neurology. In: Braunwald, E., Fauci, A.S., Kasper, D.L., Hauser, S.L., Longo, D.L., Jameson, J.L., eds. Harrison's Principles of Internal Medicine. 15th ed. Mc Graw Hill: New York. pp. 2491-2498.
- Merino, J.G. and Warach, S. (2010). Imaging of acute stroke. Nature Reviews Neurology, 6, pp.560-571.
- Swanson, R.A., Morton, M.T., Tsao-Wu, G., Savalos, R.A., Davidson, C., Sharp, F.R. (1990). A semiautomated method for measuring brain infarct volume. Journal of Cerebral Blood Flow and Metabolism, 10, pp.290-293.

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*Disclaimer: The information provided in this blog post is intended for educational and informational purposes only and should not be considered as a substitute for professional advice. While the author holds a PhD in Neuroscience and has extensive experience in the field, the content of this post may not be fully exhaustive or up-to-date, and in-text citations or a bibliography are not always provided. Readers are encouraged to consult relevant primary research articles, textbooks, and other reliable sources for comprehensive information on the topic. The author is not responsible for any errors or omissions, or for any actions taken based on the information provided in this post.*

An engaging behind-the-scenes look at the meticulous process of analysing MRI scans to explore potential stroke treatments, offering a unique perspective on the complex world of medical research.

Dr Eleni Christoforidou

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