I don’t even know how to begin writing this post. Everything that happened on a global scale in 2020 caused a lot of changes to happen in all of our lives. I will try to summarise my year…
In January, I returned to working on my project after my three-month placement at EBSOC (you can read about it here). I didn’t actually spend much time in the lab at the beginning of year because I was still analysing images from before my second knee surgery in 2019. My PhD timeline has been really disrupted and strange. So, when lockdown hit in March it wasn’t too much of a transition for me. I had been working from home a lot anyway, because my office had gotten quite busy with Honour’s and Master’s students. That’s not to say it was easy: I was feeling very anxious and found it difficult to concentrate, like I am sure many of you did too. However, I managed to finish my analysis before the summer and was then left twiddling my thumbs. The lab was closed, and any pressing work was pretty much done. I had some things on the back burner and to be honest, I was probably procrastinating doing them. I started working on my thesis, writing the sections that I could. I started thinking about my next experiments and what I would actually be able to complete within my time frame. I also taught myself how to use R (you can read about that here). I am actually pretty proud of myself for that accomplishment.
When the labs were allowed to re-open with strict guidelines and rotas, I was able to go back in and carry out two more easy experiments on plasma that I had collected a while ago.
I then set about optimising a protocol for neuronal cell culture and learning how to get it right. I wanted to know exactly what I was doing before I started the actual experiment. I will be culturing (growing) neurons from rat embryos (you can’t grow adult neurons), treating them with different hormones to see if there is direct effect of these hormones on the receptors I am studying. These receptors are called GABA receptors, because they bind the protein called GABA, which acts like the key to unlock the receptors and let other chemicals in and out of the cell. Since GABA receptors play a crucial role in a normal stress response and my previous experiments have shown that GABA receptors are affected by prenatal stress and chronic stress, I would like to know if various hormones affect the GABA receptors. This could be potential mechanism for the effects of prenatal stress. I had done cell culture before, but it had been at least four years ago, and I had never done neuronal cell culture like this. I had also never done it in the building I am working in now, so I needed to figure out where the tissue culture room even was! I spent the last few months of 2020 doing practice dissections and practice cultures until I felt comfortable with the techniques.
Turns out the dissections are really hard. Rat embryo brains are really small and really squishy. Imagine a really runny soft-boiled egg, but the size of your small fingernail. Even removing the brain is really tricky, as the embryo skulls are also very soft – about as soft as a fingernail. One I manage to get it out of the skull without squashing the brain, I then have to find the hippocampus. This is a region in the middle of the brain where a lot of learning and memory takes place. It is also an important part of the stress-response mechanism. A lot of GABA receptors that are involved in stress can be found here. I want to isolate and culture the hippocampus cells, so I have to remove only the hippocampus region. If you know what you’re looking for, you can see a small C-shaped region that is slightly whiter than the rest of the brain. And it is tiny – only a few millimetres big – so you have to use a dissecting microscope for this part. With the help of papers, videos, and a friend, I got the hang of it.
However, the learning continued and the last thing I did in 2020 was try another new technique: FISH, fluorescent in situ hybridisation. in situ hybridisation is a technique in which a tag of some kind is added to a sequence of DNA or RNA within a cell. This means you can quantify the amount of DNA or RNA in a given area. What makes in situ hybridisation really useful is that you are measuring directly in a given area, rather than mushing up the area and running PCR. I have done a lot of in situ hybridisations that have allowed me to analyse specific brain regions. The content of the brain regions was exactly the same as in life, giving me a good snapshot of what was going on. In the past, I have done all my in situ hybridisations using a radioactive tag, that creates a black spot as it decays. The method works, but it takes a lot of time because you need to wait for the radioactive substance to decay – some of my experiments took twelve weeks before I could analyse them! Plus you are working with radioactivity, which is inherently dangerous. Speaking to a colleague at work, he mentioned he used a different method: fluorescent in situ hybridisation. This technique is very similar to the one I have used, but instead of a radioactive tag, you use a protein that is labelled by an antibody carrying a fluorescent tag. You also get your results on the same day. SOLD! So, I observed the technique and learned how it works. The next steps will be creating my own labelled DNA/RNA and trying it out on my samples.
Writing it all out like these makes it seem like I did quite a bit in 2020, but it certainly doesn’t feel like it. In a PhD that has been very stop/start of the beginning, it was just another frustrating year of not being able to move at the pace I wanted to move at.
And one more thing: Covid-testing. I wasn’t sure if I should put this under work and personal. So, I guess I’ll just stick in the middle. In December, the university arranged two large testing centres for students travelling home for the holidays to get testing with a lateral flow test. I signed up to be one of the testers which meant sitting in a freezing gym hall (I am not exaggerating – I was wearing six layers and still cold) and processing swabs as they came in. I was glad to be a part of it, and happy to help, but I felt like I was cold for a whole week.
As I mentioned above, my mental health took a toll this year, but now, when I think back on the year, these highlights stand out to me:
My partner Cameron and I got engaged on my birthday in May, so that was a big happy thing to come out of a big not-so-happy year. That also meant that I could start planning our wedding and I have to admit: I am good at it. I am very good at planning things and I’ve really been enjoying it. We’ve got our venue (Jupiter Artland) and our marquee (Elegant Pitches) booked.
I gave my business Auricle a re-vamp and started seeing more sales coming in too. I have been learning so much about running a business and engaging on social media!
We adopted a third dog, Harry. This did happen in December, so it is still very new. Five-year old Harry needed a new home and we volunteered. He is a lovely little boy and very cuddly! Sid and Charlie like him too, so all is good in the land of dog.
For the last four years I have put together a community calendar of all the dogs of the local park and sold it to raise money for the Edinburgh Dog and Cat Home. From October onward I start annoying the hell out of my neighbours, friends, and fellow dog-owners to send me photos of their dogs. I also get local business to help sponsor the calendars and sell them as well. This year, the calendar did so well that we sold out within the first two weeks of it being available and had to order more! I have raised over £1000 for the Edinburgh Dog and Cat Home! I am blown away. In the past I had only ever raised between £400 and £500. I am so proud of the community and their generosity.
Weekly virtual DnD. I have been playing Dungeons and Dragons for several years now. When I started playing with my first group, we were all beginners. We had started the tradition of Thursday Night Dinners, where we took turns hosting every week for dinner and games. Soon, we started playing DnD together and learning how to play. We played through a couple campaigns until in very quick succession, people started moving away. Cameron and I started a new group, and while it was fun, it just wasn’t the same. Over the summer, we decided to try and get the gang together again for a virtual weekly session. Not everyone could make it, so we also invited some new friends to play. Now, we meet every week and play DnD using Roll20. I am the dungeon master for this game – a new experience for me and I am absolutely loving it. The group is wonderful and it is so special to get to see everyone once a week on zoom.
I think that’s enough from me for now. I hope you can reflect on 2020 and find the positives too.