This week is the second to last week of the semester before finals and everything is coming down to the wire, including my neurobio lab project. PZ was so kind as to come in and help me out this past Sunday morning; the morning after the blizzard had quieted leaving everything covered in various quantities of snow. In going over my methods we found that I wasn’t adding a drop or two of water on top of the auger layer with the immobilized zebrafish. The reason this is important is that so after the spinal cord severing is accomplished, the auger layer is separated allowing water to surround the fish immediately and preventing air exposure. The fish can then be pipetted up and put into a dish of water for observations. PZ also suggested using water with an increased concentration of calcium (14g/100mL) to facilitate better fish recovery. The fish should not be left however in the calcium water for an extended period of time because it can adversely affect development.
Repeating my methods and taking into practice the slight changes that PZ recommended, I found that after one day, four of eleven fish were still alive! After slicing up more than sixty fish with a 100% mortality rate after one day and wondering what on earth I could have been doing wrong, I was ecstatic. It’s unfortunate that this success has come so late in the game and the writeup for this project won’t show much for results other than how not to butcher zebrafish. I have learned quite a bit though about the interesting techniques I’ve been using and also about the differences in zebrafish at various stages of development. So with that, back to the lab I go to continue working with zebrafish.