11:00A – 7:50 P
- No engrailed stain
- No evidence that short fix, antibody label, then in situ, actually labels mRNA efficiently. No hb-mRNA detected.
- attempted imaging of en-dig (cut yesterday) 50 min etching removed all cells, only bits of yolk from some embryos still adherent.
- etched Ftz embryos. Not successfully stained with engrailed (see note above): nothing good to image.
- Confocal checked section and whole-mount slides. Bcd Hb hb-mRNA sections do not appear to contain substantial protein signal. Try cutting more sections. No evidence that mRNA stain worked, so try cutting samples without mRNA.
- Confocal: en-flu: weak flu signal and weak cross-reaction from green into 647 channel. Somewhat irregular brightness among some of the 50nm sections (varying thickness? or an etching thing? probably the former).
- polishing matlab dax2png and molecule list2png codes. working well.
- plan: Label with neuro-ectoderm gene battery, AP slices through densely packed embedded embryo sections, labeled with H3K4, look for H3K4me3 association with transcription.
- Review super-resolution imaging progress to date (see post)