Senior Project Week 6: Second Trial of Embedding and Sectioning

This week, I started the second round of embedding the worms for sectioning. This time, we will be cutting a full worm into 5 parts and a full isolated worm nerve into 8 parts. This is in order to give us insight into how the nerve chord narrows as you travel in the posterior direction in order to evaluate the best places to stimulate and to record for the next round of stimulation. We preserved the samples in Eppendorf’s filled with Paraformaldehyde, or PFA, and then proceeded to dehydrate the samples along with another batch of human cochlea. The process is as follows: 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. We will then slice them in 10 micron slices by the ultratome in order to gain cross sections to analyze the width changes along the course of the nerve from caudal to cranial.