Thursday 07/5/12

9:50A – 7:15P, 9:00P – 10:45P

  • check nascent transcripts of AbdA (red) and Antp (green) on yw and Pc O/N short-fix embryos.  No obvious staining on turnkey.
  • Should check 6/11 embryos as positive controls and 6/25 embryos to see effect of long-term methanol storage:
  • Probe test transcripts clearly visible.  Further inspection 6/25 slides do have decent transcripts: also confirmed in sections.   Substantially less bright than probe tests, background substantially higher.  Might be post-fix GA treatment adds background.  Might also be substantial antibody loss.
  • Whole embryos (after postfix?) from 6/25 do show clear nuclei detectable in 647 from H3K27.  However sections don’t — nuclei are primarily voids.  Very different from previous images of directly labeled K27 with Psc (see lab notebook images).  Need to increase the K27 antibody concentration.
  • Organizing references for snail bistability paper / intro.
  • Fixed all duplicates in Mendeley Library
  • sectioning yw Su(Z)12 H3 AbdA/Antp stained embryos, demonstrating for KE how to pick up sections on coverglass (should solve substantially non-flat problem.  Might change etching rates?
  • STORM imaging of Antp H3 and K27 from 6/11.  K27 stain too weak suspect problem with antibody. H3 somewhat weak, probably buffer run-down.
  • Finish embedding DNA FISH embryos.  At 70C, 10:00 PM
  • Flip collection cages 10:15 PM
  • Revisions on dynamics paper back from ML.  Need minor edits and format changes, check against Nature Methods guidelines.
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