Laser Powers:
- 750: 100 mW
- 647: 15o mW
- 561: 150 mW
- 488: 15o mW
sample ‘4H’ (histone H3 stained in all 4 channels as an alignment control).
- etched sample 22 min. Looks like all resin is dissolved.
- very weak switching. Either new glox is bad. Or there is still too much resin in the tissue.
Imaging slide S2: (engrailed).
- slide is 1 wk old, H3 switching does look less (20K frames of good switching instead of 80+).
- 750 switching even more sparsely.
- Setup multiposition run. Had 561 ‘run shutters on’ = no data and 647 laser off = no data. Damn it. Fixed after movie 6.
Imaging slide Psc-hox. Only imaging 647 and 750 channel. No nascent transcripts in 561 ID in overview scans (even at 100x).
Etched new slide of H2. 20 min, +5 min + 7 min .
- Etching still seems to have failed: after 2 hrs arraying slide, attempt STORM calibration tissue glows brightly under high laser, washes out single-molecule signal.