Friday 10/19/12

9:30A – 5:00PM

Embryo fixation

  • 9:30A Flip collection plate
  • Making new buffer A fix mix 10 mL, fresh.  (Should have made 100 mL and froze aliquots?)
  • 10:30A Fix embryos.  Now in -20C in MeOH.
  • Write to lab and Francis lab about starting S2 cell culture.


  • remove Pc antibody/embryos from 4C for 1hr RT incubation.
  • remove and store Pc antibody for reuse.
  • Rinse out antibody (1hr+)
  • post-fix (heat-reactivate post-fix mix at 60C), 5% form 1/2x PBT, 25 min.
  • rinse out post fix.
  • Dehydrate into MeOH and store at -20C

Data analysis

  • computing per cell cluster properties (more data points than per-image).
  • As suspected, not a very robust measure.  sharp segmentation of cells is difficult, and more numerous less robustless estimated small cells (nuclear fragments) dominate the analysis.
  • Basic trends still captured by analysis though.
  • Organizing results in ppt presentation for Francis group meeting
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