9:45 A – 12:45 A
- Evaluating 3D fitting: section joins do not look smooth
- Wrote new script (list2tif3d_weq.m) which distributes samples uniformly in z. Samples near the focal plane are best resolved and can have higher z-resolution.
- Current approach still involves sacrificing molecules in last two bins. At expectation of 400 nm reliably returns 280-290nm sections, which rejects about 60% of total dots found
- GFP flies arrived today
- Etching rad-21 and 2H sample at 50 rpm agitation for 50 min.
- Raw squares of resin still visible on rad21. Etching and agitating for another 24 min.
- Attempting spline registration to improve 3D reconstruction of mitotic cells.
- STORM imaging of samples:
- 2H
- rad-21
- O/N run on sample H2 (H3ac-750, H3-647, H3K27me3-561). 3 chn only (488 looks blank).
- on all samples: 750 working well, 647 and 561 resin questionable?
- m a-dig antibody arrived
- Restart Bcd, Hb, hb-y-(i) staining.
- Also stain Psc and Pho labeled embryos with PcGs in 750, and Dmo1 in 488 (will put K27 in A647 channel and H3 in Cy3B).
- see records, slide 101 and 102: