Laser powers
- 750: fix to 100 mW
- 647: 175 mW
- 561 (at 200 mW activation) fix to 150 mW
- 488: 200 mW
- 405: 1.5 mW
Imaged S6 500 nm, single section with hox expression dot.
Arraying sample of S7, engrailed expression with polycomb staining in 100 nm sections. engrailed expression clearly visible in 10x overview images.
At movie 30 (probably happened earlier) found z-focus way off. Unlock and relocking the focus lock restored accurate focus. It seems to me this shouldn’t happen… Probably occured during large spatial jump from bottom of to top of section. This will be an issue for imaging multiple ribbons on the same slide.
Etched S2-engrailed 330 nm sections for 35 min. center of embryos fully dissolved, will see if this is over-etched.
O/N imaging of Psc-en. Good 750 imaging.
Etched Psc-hox (330 nm sections) for 25 min. Some segregated bands that still look like resin, but embryos look liberated, possibly over-etched possibly under-etched. did not get fluorescence illumination working right on Turnkey to check (had only 30 seconds to check since Turnkey scope was in use). Mounted both samples for STORM viewing.