10:05 A – 12:10A
- PcG and Bcd staining embryos incubating in primary at RT for final hour.
- Flip fly plates
- Discussion with John about projects
- Need to look up Drosophila repeat sequences for probe testing.
- in situs day 1 on PcG, Bcd and 4x snail embryos.
- Some primary labeled, post-fixed PcG and Bcd embryos put aside in methanol and stored at -20C. The antibodies should survive as well as any endogenous protein in methanol at -20C for long-term storage.
- Confocal of rad21 – looks good: uniform nuclear staining.
- Confocal of engrailed mature and nascent mRNA detectable in S7 both in resin and in STORM mounted slides. Try again on STORM scope on Fri?
- Starting work on Fly meeting talk.
- Flip fly cages.
- Sectioning: 3 slides of Psc + hox + H3.
- 3 slides of H2 at 330 nm
- 2 slides of S2 (engrailed + H3 + H3K27) at 330 nm. 1 slide at 250 nm.
- Slides in the evening (H2 and S2) not flattening out nearly as much with chlorform as Psc. Resin reacted longer and has been dry for longer…
- Plan to do 3D bead calibrations on STORM2 with Yari tomorrow.
- in situs incubating O/N at 55C. Added fresh dose of hox-dig probes to previous mix. Made fresh batch of en/inv-bio, E(Pc)/tou-dig, fresh y(i)-dig and fresh sna-dig sim(2A)-bio.