10:00 A – 7:40P, 8:10P – 11:30P
- Meeting with Manching, discuss new strategies for single tagged histone viewing. (see email)
- Preparing bead slide for calibration of Confocal images, 1:5000 and 1:10,000 dilution in PBS w/ Mg and Ca. Flow into slide/cleaned-coverslip chamber made with doublestick tape. Rinse several times with PBS w/o Mg or Ca. Then seal.
- Fluid flows faster around edges next to double stick tape.
- nicked translation with unlabeled dNTPs
- Making tetraspec beads at different concentrations — high concentration works best (3x dilution). Imaging on Zeiss700.
- Imaging MP08 snaD on Zeiss710. No correct age, correct orientation, correct genotype embryos. (maybe 200 embryos on slide, many non-fertile too/young).
- Imaging MP07 snaD 30C embryos on Zeiss710. Only 1 good embryo, many many too young.
- Laminin antibody definitely stains cell membrane in early embryos. Does not surround nuclei in pre-cycle 14 embryos. — basement membrane factor, useful for cell tracing, not nuclear factor.
- Nicked translation from plasmid directly does not work.
- Running Zeiss710 ON, doesn’t seem to have center and offset for z? using fixed first and last — (2 sections of same embryo) hopefully this works. Check tomorrow.