Rat brain
Week 5 July 13- July 17, 2015
This week was full of entertaining activities, with perfusions being the most interesting thing. The first few days consisted of preparing formaldehyde that would be used on the CRF rats, under Giordano’s research; helping Juan, an undergraduate student, move the rats and clean their cages; and watch drunk rats try to maneuver after they passed out.
Research requires a lot of tedious work, with long hours of waiting and watching in order to collect a few snippets of data. On Tuesday Molly, Giordano, Kole, and I took turns watching eight rats wake up after they were administered 4 ml/g of ethanol that caused them to pass out. This is called a loss of righting reflex(LORR) test.
After they were injected with the substance and fell asleep, the rats were placed on their backs between triangular prisms. The first four rats were asleep for about four and a half hours until they finally woke up and turned over. The test required that, in order to officially declare them awake, the rats had to be able to turn over three times onto their stomachs (we turned them back over onto their backs once they succeeded) in a minute and then be able to turn over three times, three times in a minute. The chaotic part was that they all started turning over pretty much at the same time so it was tricky to figure out how many times each rat had been turned over so far. Nonetheless, we were able to successfully do it.
Once the rats were officially awake, Giordano nipped their tail and drew a couple drops of blood into small, separate containers. The other four wild rats that had yet to wake up didn’t actually do so until 7:00 PM. Our mentor Giordano had stayed with them. The research these rats are a part of is a collaboration with another lab.
We also aided Molly in mixing a 4 percent formaldehyde substance. Because formaldehyde is toxic we were required to wear gloves, eye protection, and masks. It felt very scientific. The procedure was relatively quick and the mixtures we made were acceptable. This solution was then used on Friday and injected into the rats that were euthanized to result in tissue (and brain) fixation. This allows the brain to be cleanly cut into super thin slices, with little mess and preciseness so that the brain can be well observed and data collected from it.
On Friday we aided in the perfusions of the rats. We were assigned the simple tasks of getting the right rats from their cages, gathering the needed materials, preparing the materials, carrying the rats, and chopping off their heads.
When we first bring the rats in they are put into a chamber that has a supply of CO2 being filtered in and causes the rats to lose consciousness and eventually die. To better preserve the brain, the formaldehyde solution is injected into the bloodstream and thus out to the rest of the body. After a bit Molly takes the deceased rat and extracts the brain. Most of the brains ended up pretty perfect with few errors or mistakes being made. Once out of the skull they were inserted into glass bottles with formaldehyde and put on ice. This whole process was amazing to watch.
I also helped out Marsida in filling out the lab book for the 11 rats in the self administration chambers, who had all been sacrificed one per day a while ago. It mostly consisted of copying numbers into data tables and erasing pencil marks. Nonetheless, it wasn’t a bad way to start off a Friday morning. And going into the procedure and doing the perfusions made Friday a good end to the second to last week here at Scripps. Looking back at the past few weeks, the time has really flown by but somehow been able to cram a lot more knowledge and experiences into the hours working at Scripps. I have greatly enjoyed this experience working here so far.
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