I have crazy intense mixed feelings about weekend science. On the one hand, it is an opportunity to make a lot more progress. On the other hand, it takes away from the little time I get with my son. And nothing is worse than devoting the weekend to science just for nothing to work. It feels like such a waste.
This weekend was not like that. I have never before had such a successful day of science, especially over the weekend. I have been working on learning a very difficult experiment, one that has so many opportunities to fail. This Saturday, everything fell into place. Here is everything that had to go right:
1. Mouse had to get pregnant and stay pregnant and be at the correct stage of pregnancy when I perform surgery.
2. Surgery on the mouse: successfully extract embryos, inject DNA into the small embryonic brain region, electroporate the DNA into the front cortex, place the embryos back into the mother, and suture her up before she overdoses on anesthesia.
3. Mouse to stay pregnant for three days after surgery and embryos survive.
4. Embryonic brains have to show fluorescence (from the DNA) in the correct region of the brain.
5. Successfully dissect and flat mount the cortex.
6. Genotype the embryos and have a positive band for the correct mutation (theoretically only 50% of the litter will carry it but it can be more or less).
7. The flat mounted cortices have to be flat enough for the microscope to be able to image it.
Only when all these things go right (over the course of 2 1/2 weeks) to collect one sample set. If I’m lucky, I’ll get at least 5 cells undergoing division to quantify from this sample set. I need to collect about 20 cells undergoing division. But to have the level of success I saw yesterday (all the embryos survived and 3 of them had fluroescence) was a miracle to top all miracles! I needed this so bad, I needed to be able to show my boss that I have the ability to learn and succeed. I will never enjoy losing a day with my son for science, but at least it was worth it yesterday.
