9:00 am – 5:00 pm, 7:45 pm – 10:45 pm

practice talk (9am-10am)

Lab meeting 10am – 1:30pm

• see protected notes

Embryo MERFISH project

Embryo staining

STORM3 observations of cut sections

• some individual spots look promising.
• despite high concentration of probe, background does not look any higher / not too bad (still pretty clear to see tissue, no mistaking cytoplasm with slide background. Cytoplasm background substantially higher than nuclear background.

confocal observations

• some staining of tsl, not substantially better than samples from yesterday stained at lower concentration in 35% formamide.
• sample quality I think is compromised — some nuclear morphology looks poor, DAPI channel background in the cytoplasm looks unexpectedly high, and nuclei are not as bright as I would expect for a substantial, long incubation in more concentrated DAPI (1:100 dilution instead of 1:5000 of what I believe is a 5 mg/mL stock).
• conclusion: we can get some staining. Further diagnosis will be safer with fresher samples. Let’s cut fresh embryos. Compromised sample integrity may have been a problem in previous chromatin stains as well.

Fly work

• transferred both wildtype tube adults to new bottles. By the time I get back from Genome Biology meeting there should be plenty of these.
• if GS joins the lab we can start working from new embryos together.