Thursday 04/23/15

9:00 am – 10:15 pm

Microscope booking

  • booked STORM3 for imaging this evening
  • STORM2 in use, Andor camera moved to STORM5 construction scope
  • STORM4 may be available for checking samples
  • re-applied for expense code for confocals.
  • registered for 7pm – 8pm to for confocal LSM 700 to check embryos

Embryo staining

  • embryo sections on coverglass on elevated platforms dried out :(. Should seal with rubber-cement or cover with a wet kimwipe. humidity chamber not working too well.
  • no staining detected in Tll sections
  • check embryos on Turnkey? — No camera
  • check embryo staining on LSM 700 — no staining detected in blastoderm embryos. Late embryos seem to have some staining around the CNS and around the

Staining try 2

  • 10 uL in 100 uL of hybe dilution buffer made with 10% dextran sulfate and 10% formamide.
  • not using spacer on slides this time, 20 uL of staining buffer, sealed with rubber cement.
  • added embryos (~15 uL) to remaining 80 uL of hybe dilution buffer with 10 concentrated probe
  • (this time its clearly blue, so probe concentration is super high). I expect we get background and I can work from here washing it down. I hope we get decent signal too though. There didn’t even seem to be good single molecule background signal when tested on STORM4 today.

more problems

  • something went wrong in probe assembly, probes sequences for tll map to tsl:
  • this looks something like the confocal pattern today (From BDGP):
    tsl_staining
  • and my ‘Kr’ probe is mapping to lea, (aka robo2), no pattern recorded in BDGP. should have checked these earlier.
  • found bug in code: GeneSelection_150325. gene names got mismatched with sequences during select longest isoform phase…
  • need more investigation tomorrow…

Oligopaints team meeting

  • at Wu lab.
  • good presentations,
  • hierarchical scales of coding from Sonny look good
  • resolution depth and speed from Peng Yin lab look pretty awesome.
This entry was posted in Summaries. Bookmark the permalink.