9:30 A – 8:10 P, 9:00 P – 12:10 A
Install cryo-attachment
- be sure to retract arm and center FC mount prior to mounting
- be sure there is space between the arm and the chamber so the arm can move
- remove chuck before baking system at end of sectioning
Things to get / do
- cryo-loops (Yari has some)
- knives + glass knives for training (get from HMS)
- Oxygen sensor
- Contact facilities about humidity control (Colenso will be point person)
- Contact facilities about oxygen
Cell labeling
- With Ajaz:
- staining Ph-FLAG, Ph-mutant-FLAG and S2 cells (neg control) with 647 anti-FLAG
- 15 min in 4% PFA fix
- 7 min in 1 mg/mL NaBrH4
- 3 hr in primary anti-flag at 1:200
- 1 hr in secondary R&D 2:2 anti-m A647, A405
- set up STORM imaging of Ph-FLAG cells O/N
STORM
- Finish O/N imaging of 405,cy5 Fab7 (ran out of diskspace. 512×512 eats disk space fast, >1Tb night (instead of 250-350GB)).
- 900 mW 647 laser
- O/N imaging 512×512 of Ph-FLAG cells
Review
Stuff to do
- book hotel for SDB