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Aaron Uy - Week 6 and 7


Week 6 and 7

These two weeks were really busy. Sarah - an undergraduate - and I continued on the IHC project and I perfused 3 more mice so we could do IHC on them. I’m getting better and better at perfusions (and more desensitized to the gore). Though, I always remember to say a little prayer for the mice before I cut it open and thank it for its scientific contribution. Hopefully they are all doing fine in mice heaven. Because mice brains are sliced and sectioned off into 6 wells, we were able to perform IHC on 6 batches of brain slices, 2 from each mice, (the other 4 batches are preserved and saved in case they need to be used in the future). Of these two batches, one was stained for the somatostatin receptor, and the other one was stained for the parvalbumin receptor. After IHC, we mounted the slices and then viewed them under a microscope. Unfortunately, the slices were a little beat up, but overall the entire staining went well.

During this process, I continued on my semi-automatic-analysis code in ImageJ. Having it run over an entire folder of images and then saving to a specified directory was a lot harder than I initially expected. I wound up rewriting a large portion of it and in doing so ran into a lot of troubleshooting errors. These issues were furthered by the fact that I don’t have much coding experience. Thankfully, an undergraduate there, Jack, was able to help me with the few things he knew about the ImageJ coding language. Eventually, I finished and was able to successfully run it over all the images of the slices we performed IHC on, effectively cutting a job that would have required days of monotonous labor of counting the cells into something that could be done in an hour unattended. My PI told me to write up a protocol so that the lab could use when I’m gone, and so I started on that.

Also, I assisted a ton with my postdoc’s mice surgeries. In all we did about 20 over a few days. These cranial injections involved injecting a virus into certain locations in the brain. They then could see effects of the inhibition of certain parts of the brain during behavior tests and deduce the importance of certain locations and structures. Initially, my role was to find the coordinates of the injection site and inject the virus. Eventually, I was able to perform my own surgery, exempting a few parts, but for the most part it was my role to cut the skin and tissue, align the coordinate values, drill holes, inject the virus and suture the skin.

Meanwhile, I continued to help with the behavior tests my PI was running, attended a talk that one of the other postdocs gave, got to know the lab members more, ate a lot of yummy food, and continued to enjoy my time at the Fuccillo Lab.

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