It’s already week 9! Welcome back to my blog!
This week I continued
watching videos for Day 7 in cages 1, 3 and 5 and have made my way through a good chunk of that footage already. Although I have not finished analyzing the data and comparing it to the Day 1 and Day 2 data from the same cages, I did note one weird aspect in Day 7's behavior. Unlike in Days 1 and 2, where the majority of aggressive behaviors fell under the umbrella of posturing and scuffling, in the Day 7 videos, the majority of recorded behaviors so far are biting. I believe this is due to the fact that the mice have already set up an existing hierarchy in which the dominant mouse bites the others to "remind" them of who's in charge. This is very different from the subtle aggression that we were accustomed to seeing while the mice were still creating their cage hierarchies. But I will update next week depending on what the rest of Day 7 data shows!
One of the coolest things I did this week was the LPA Immunohistochemistry test I ran with
Bret over the last three days! An extremely involved staining process, the LPA
IHC test spans over nearly five days and requires time and humidity sensitive
steps that must be done with extreme attention to detail.
We decided to run a LPA IHC based on a very specific procedure and ran an optimization test to determine the best concentrations of blocking (base) solutions to use to hopefully increase the contrast and quality in the traumatic brain injury fresh frozen brain tissue.
The tissue that we are examining belongs to two different animals, one that was injured as a part of a traumatic brain injury (TBI) test and the other that is a sham animal. A sham animal is one that receives surgery and treatment that the injured animal receives without actually being injured. The purpose of the sham animal is to isolate the effects of the injury in the TBI animal by using the sham as a control for the effects of the surgery and/or treatment. Both brains were fresh frozen, which means they were not treated with paraformaldehyde after being extracted and can begin to decay when they are allowed to thaw.
Although I am not an expert in this technique yet, it was very interesting and different from most of the work I have done in the lab so far. Although I worked alongside Bret for the majority of the steps, I mixed and applied the control primary antibody solution while Bret completed the primary antibody application and completed the majority of the rinse and drying steps by myself.
We decided to run a LPA IHC based on a very specific procedure and ran an optimization test to determine the best concentrations of blocking (base) solutions to use to hopefully increase the contrast and quality in the traumatic brain injury fresh frozen brain tissue.
The tissue that we are examining belongs to two different animals, one that was injured as a part of a traumatic brain injury (TBI) test and the other that is a sham animal. A sham animal is one that receives surgery and treatment that the injured animal receives without actually being injured. The purpose of the sham animal is to isolate the effects of the injury in the TBI animal by using the sham as a control for the effects of the surgery and/or treatment. Both brains were fresh frozen, which means they were not treated with paraformaldehyde after being extracted and can begin to decay when they are allowed to thaw.
Although I am not an expert in this technique yet, it was very interesting and different from most of the work I have done in the lab so far. Although I worked alongside Bret for the majority of the steps, I mixed and applied the control primary antibody solution while Bret completed the primary antibody application and completed the majority of the rinse and drying steps by myself.
Because of the nature of the drying process of these slides, the
results of our stain won’t be available until early next week, but I’ll be sure
to include the results and pictures of the slides we stained in my Week 10 blog!
Thanks for reading!
Tasha
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