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Mice's Brains and Western Blots

One of our co-founders, Megan's, experience at a laboratory at the University of Hong Kong.

One of the reasons I co-founded BecauseMentalHealth was because, among of many of the friends and colleagues I've talked to, almost none of them knew what neuroscience was, much less neurodegenerative diseases, but I was always incredibly fascinated about it, so when I found out my application as a Research Assistant/Intern at the Lab of Neurodegenerative Diseases in The University of Hong Kong was accepted, I was ecstatic.

Originally wanting to intern during my Winter Break, it was postponed to the summer due to COVID and other unforeseen circumstances, but I was happy to do it over my Summer Break since it meant I was going to work for a longer time.

My first day at the Lab was Monday, July 5th, 2021. I went to the lab in the afternoon, got shown around and introduced myself to my research group, which were all working on the relationship between periodontitis and Alzheimer’s disease. Then, we were sent to a Lecture Theatre to listen to the lab's professor, Dr. Raymond Chang's, beginner lecture series on neurodegenerative diseases, which he graciously offered to the summer students, stating that this year's batch was larger than usual.

On my second day, the Thursday of that week, I finally rolled up my sleeves (figuratively, of course- we wore lab coats and gloves to protect ourselves as well as our samples) and got to work. I worked alongside two of the lab members in doing Protein Assay to prepare for the Western Blot the next week.

Since our school had less of a hands-on approach to learning, my experience with laboratory equipment was little to none, so when I had to use professional equipment, my heart raced. Even using a simple microliter pipette was something new to me- even making mistakes (thankfully, small ones that were reversible and did not impact our results) that caused me to internally shudder and cringe. But thankfully, after a few demonstrations by the other members, I quickly picked it up.

The next few days at the lab were used for Western Blotting and the procedures preceding and following it.

Before Western Blotting, we had to prepare the samples, which consisted of many procedures- the last of which was reducing and denaturing the samples by boiling the proteins.

And here came to a good part- Western Blotting- which is a form of gel electrophoresis.

First, we had to make gels, which was both an excruciating yet fascinating procedure. Then, there was the electrophoresis buffer (aka Running Buffer), which provides ions for the passage of electricity; the Stacking gels, which are used to keep the protein samples in line - though they were made of the same ingredients, they contained different amounts of each. Since they were supposed to be a jelly-like structure, we marvelled for long times over how the tubes we kept the remaining mixture could be flipped upside down but nothing would spill out.

The samples were loaded into the machine, and then they would slowly travel down it, according to the weight of them- the heavier they were, the slower they travelled.

After the Western Blot was finished, we had to transfer the proteins from the gel to a membrane for future analysis. Putting the membrane in a large sandwich of saline-dipped filter paper, sponges and right next to the gel, it was then placed in another machine, and the process was complete.

And after transferring, we did imaging to analyse just what genetic components were in the samples. It involved light-sensitive chemicals and some lengthy periods of waiting, but all in all, it was quite an interesting process.

During my last two weeks of laboratory work (August 9-13 and 23-27), we solely did animal work, which I could not take any pictures of due to Animal Ethics Laws. However, it was truly an amazing and fulfilling experience.

The two behavioural experiments we ran were the Spontaneous Y Maze and the Puzzle Maze tests. The Y Maze test consisted of analysing the data of the mice going through a maze shaped as a Y, and the Puzzle Maze consisted of watching the mice slowly go through an easy maze, testing their problem-solving skills, short-term memory and long-term memory.

Then, after the behavioural tests were done, it was time to sacrifice the mice for their samples to analyse. It all took place in a room with no windows, filled with cages (mostly from our group) that contained mice excretion, which did cause us to need to hold our breaths every time we walked near them. There were many samples we needed to take- liver samples to analyse for systemic inflammation, gum samples to analyse periodontitis-induced inflammation, and brain samples (in the regions of the hypothalamus, the frontal cortex and the hippocampus).

There were many different uses for the samples- DNA Extraction, Western Blotting and Imaging- and we were taught the many different ways to get the samples for them. Since Imaging required a clearer sample, the mice were perfused with saline, flushing all the blood out of their bodies in order for the process to be easier and the Imaging to be clearer. And though my parents were disgusted as I explained all of my experiences with them, I thought it was truly valuable and helpful. I mean, hey, at least I know the basic structure of a mouse's brain and how to perfuse them now!

Before this experience, I didn't know exactly what Alzheimer's was caused by, and how Periodontitis and inflammation was connected to it, and I certainly was not familiar with many laboratory techniques, but all this knowledge was instilled into me in merely two months. But most of all, I made four amazing friends that were caring, enthusiastic about learning, and happy to help me (which I needed a lot.)

I highly recommend you try doing some experience in the field you are looking to specialise in, not only to learn new things, but to find out if that path is truly for you. I’ve found that I’m still very passionate about the brain, especially regarding Alzheimer’s; but perhaps research is not for me since I like more hands-on, talking work.


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