10:30 A – 6:20P, 9:30P
Embryo collection
- Flip all fly cages
- dispose of previous fly plates with new plate recycling and freezing system.
- Fix embryos 9:30 PM (~3-14 hrs, mostly at raised at 29C).
STORM
- working out parameters for TCEP imaging
Current working set:
- 128×128 field of view.
- 310 mW 647.
- 1750 mA of 752.
- 2 mW max 405.
- 60Hz frame rate for both. 45K frames of 750 for BXC. maybe 60K frames for Cy5 BXC.
- 14.2 TIRF (any higher 750 fails. 647 would definetely prefer a slightly higher TIRF angle).
- should have had longer bleach fro 647. (surprisingly little blinking for the amount of bleach time (~7000 frames at 60Hz).
observations
- signal to noise actually looks much better in 750. The slide may have aged a bit poorly, or this buffer is less good for cy5.
Experiment planning
- embryo staining tomorrow, image on Wed.
- Order primers for rest of LC library for chromatin colors
- emPCR of rest of LC library on Thursday (starting at noon).
- Fab7 start biological replicates in two weeks (Brian will make more probe after his department retreat).