10A – 11:45A, 2:ooP-4:00P, 6:00P-11:50P
- Turnkey 60x works alright for mitotic sorting of DAPI staged embryos — constructed large well coverslip with channels for removing excess buffer. In mininmal buffer embryos don’t float around and are easier to pick off of coverglass with a brush.
- Currently segregating embryos in chambered slides which allow for rinsing and viewing at high mag.
- Select 4 mitotic embryos for Yari and Graham.
- 20x air on Turnkey insufficient for mitotic staging — difficult for cell cycle staging.
- H3K27 stain works well — strongly visible in late (post-germ band extension) embryos. Hardly detectable in cc14 embryos. Not at all detectable prior to cc14 (based on bulk nuclear fluorescence)
- Resin embedding embryos stained with H3K27 in 8 chambered slides. Using disecting scope and pipette (with back-up barrier for resin) to transfer embryos between washes. 20 uL easily sufficient to pick up embryos, should try cutoff P10 tip.
- Finish 2ndary rinses and post-fix primary ON embryos.
- Primary ON embryos in PBT on bench, unused rapid primary embryos in drawer at 4C (also in PBT, already post-fixed).