Friday 03/29/13

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

  • working on cooperativity section notes_on_kolmogorov

Stuff to do

  • book hotel for SDB
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