Thursday 02/23/12

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.
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