9:40 A – 7:40, 9:00P – 12:30A
- Finished confocal run, 8.5 embryos finished at speed 5 by 10A (run started at 10P).
- Previous user canceled 710 time (again). Need to check before starting or stopping a run.
- Over-etched first K27 slide (17 min + ? 15).
- 750 laser on STORM 4: Cathode I low error on boot up. Automatically shuts down.
- Prepped new bead slide 715 and 540 beads at 1:10,000. # prep bead slide of just 540 beads.
- Making fresh amp plates
- Attempting to troubleshoot vpn connection to Cajal and Monet with IT help.
- DNA FISH labeling of embryo sections, day 2. Sections still survive. Not sure if nuclei are intact, most obvious signal appears to be yolk vesicles.
- Confocal imaging slide m199. Most embryos appear strong rescue.
- Attempting to run BowTie on a subset of K27me3 data in Galaxy.
- 750 laser boots correctly after turning off main power box and letting cool.
- STORM imaging of K27-control. 750 a-m (Dm0) and 750 a-rb (K27) either out-competed by the 561 and 647 antibodies, or the dye doesn’t work, or the over-etching killed it.
- in situs day 3 (KE) 3 color hox
- [spreadsheet 0AjSqkxgziU1YdG90a29UaU5ua1FLUXZ5LTFmM1pMSlE 602 202 sheet=8]
- Confocal imaging of 3 color Hox. 750 didn’t work (no 405 nuclei detectable). Most channels look good.
- # Test if etching interferes with dyes
- # Try DNA FISH with embryos on slides instead of PCR tubes?