Day 16

Ink Microinjections
In order to determine whether the microinjections given to the rats throughout the experiment were isolated to both sides of the parabrachial nucleus, we give rats microinjections of dye.  Whoever is doing histology will then cut the brain into pieces, put them on slides, and examine them under a microscope to see where the dye is located.  Our goal is for the dye to only be in the PBN (bilateral).  Lindy let me give the rat the ink microinjections.  First, they are injected with anesthesia.. Lindy let me wrap the rat for its injection as well, which was a lot more difficult than I expected.  Every time I correctly positioned the rat, he would move.  Sometimes he would even grab the cloth and unfold it when I picked him back up to try again.  The time I finally got him wrapped properly, it was not tight enough and he could still move when the needle was being injected.  Lindy eventually had to go ahead and do it herself.  We then waited for the rat to go unconscious.  I unscrewed its dummies and placed the injectors in the cannulae.  I also set up for microinjections.  The hardest part was getting the tubing onto the microinjector without having air bubbles.  There would be air bubbles where the microinjector inserted into the tube, and  I poked a hole in the tubing a couple times.  It took a few tries to finally get it right.  Once the microinjections were complete, I would hand the rat with its label to Dr. Holstein for perfusions.

Cleaning
Julia and I deep cleaned the testing room.  We took apart the testing boxes and wiped down all the solution that had stuck to the counters and floor.  This is important so the mess does not accumulate over time.