Senior Project Week 4.5- Embedding

I am writing this on the week after spring break, but, being the diligent researcher I am, I still performed lots of research over spring break. Last week, me, Jen, and Claus spent a lot of time prepping and performing the embedding and microscopic sectioning for the samples. The worms intestines were removed in order to verify the presence of the ventral nerve chord in the anatomically correct spot. Next, we had whole worms dehydrated and embedded with the following process- 25% acetone 15 minutes, 50% acetone 15 minutes, 75% acetone 15 minutes, 90% acetone 15 minutes, 100% pure acetone 15 minutes, 100% pure acetone 20 minutes, 1:7 plastic to acetone 1 hour, 1:1 1 hour, 7:1 1 hour, pure plastic 1 hour. They were then sliced in 10 micron slices by the ultratome in order to gain cross sections to locate the ventral nerve chord, and location was performed under Phase Contrast, Darkfield, Brightfield, and DIC. Next in the process is segments of the nerve chord itself, which was dissected out by cutting again on the dorsal side, removing intestines, pinning down top skin to the side, and cutting off excess skin around the nerve chord, followed by sectioning the isolated nerve chord into 3 sections and preserving them in formaldehyde. They will soon be embedded and examined in the 10 micron cross sections. Next week will also contain some stimulation and the first recordings of our CAPs.