10:40 A – 12:30 A
- PBT rinse for embryos, move to block
- stored block is cloudy at top again. Make new block with filter sterlized PBS, add new tween 20 separately (150 uL in 15 mL).
- Bike to MGH to borrow small sample of antibodies from Manching
- Chemistry Lab Safety training 2p-3p
- Sectioning time on ultratome 3p-7p: cut many good ribbons of H3K27-647, en-555, WGA-488 stained embryo.
- Confirmation of sectioning on Zeiss700, 4p-6p
- New format for documenting embryo staining.
- Testing new invitrogen DNA dyes (8 in total). Incubate at 1:500 1 hr. wash out, incubate in PBT O/N at 4C.
- Test en-555, WGA-488, H3K27me3-647 stained sections on Zeiss700 — staining looks good, (except 647 as before can’t see staining in embedded sections. Can only see 488 staining under oil immersion). go for SuperResolution imaging.
- New antibodies testing, O/N secondaries at 4C
- Filter chambers need more than 500 uL to avoid fluid getting trapped only in upper part of chamber.
- bcd hb staining at 1:100 O/N at RT in 15 min GA PFA fixed embryos.