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):
- 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.