10:00 A – 7:40 P, 8:30 – 10:00 P
General
- laboratory safety training refresher
- check HoxD server jobs: still heavily backed up
- check STORM analysis / drift correction: Y complete. Others still running.
- update notebook entries from yesterday.
Review
- working on rewriting introduction
Embryo staining
- Select and image En-embryos (try 3)
- Staining two coverslips of En-embryos with 10 WGA diluted 10:500. Staining looks great.
- using 4x objective to array entire coverslip (try to avoid the ‘unable to relocate embryo’ issue from last time!)
- Image 561-En stain of all embryos in all patches
- coverslip 1, 40 min PBS-Tr (.5%) treatment, 8 min HCl (.1M) treatment, then prep for hybe.
- coverslip 1: En primary unlabeled batch 1, 5 uL + secondary-cy5 with docksite 1 uL + new 405 tertiary (no overhang), 1.5 uL in 25 uL hybe.
- coverslip 2: PBS rinse, PBS-Tr 10 min, then pre-hybe at RT
- coverslip 2: En primary labeled (batch 2), 6 uL in 20 uL hybe.
Ph project
- STORM of PhM-flag-488 + Ph-wt-647