Tuesday 07/03/12

10:00 A – 7:10P, 8:45P-11:00P

  • Sent version 4 of snail dynamics manuscript to ML
  • Flip collection cages.
  • 12:30 PM fix Pc-GFP and yw embryos.  15 min fix.  Rehydrating embryos to test protein stains.
  • Image sections on Turnkey.  sections substantially non-flat.  more chloroform / drain bath slower?  Might need to demo a few more slides for KE to improve her sectioning + collection.  Ribbons and embryo density look nice.
  • In situs on embryo stock collection O/Ns do not look good — some detection of of Tub.  intronic mRNAs not readily detectable, dramatically worse than 6/11 in situs.
  • Teaching bead calibration on STORM2 to KE.
  • KE STORM imaging of slide 4 (AbdA, AbdB) from 6/11 batch.
  • IR focus lock not working.  Imaging session aborted.  Talk to Hazen tomorrow.
  • DNA hybe following Bantignies protocol.
  • important change: probes denatured for 15 min, embryos for 10 min, then added together, dropped to 37, mixed by pipette, and denatured another 10 min at 80C.  Next time try 15 min at 83 for embryos, mix by pipette. 10 min at 95 for probe, add with both hot for 2 min, then 37C 17 hrs.
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