9:30a – 8:30p
issues with z-calibration
- fixed bug missing
sqrt()inwx = w./sqrt(ax)andwx = w./sqrt(ax) - take zcal bead images. Run Z-calibration script on these to generate curve fits
- use the curve fits to refit the beads, plot the reported zs. Should be by design mapping to 0, jump, linear ramp -600 to 600, drop to 0.
- This is not what we observe: still doesn’t have the correct z-range. Off by a factor of 2.
Microscope meeting
- add dual view to STORM3 (me?)
issues with 750 STORM
- yesterday, detected no 750 staining on sample G6-G8 previously imaged beautifully in 750 over the weekend (when the 647 laser was off!)
- today tried probe hot wash in water-bath at 60C on heat block pre-heated to 60C. Measured temperature in deep filled well also reached 60C. Sample may have been up to ~65C.
- no 750 spots detected, just very few stray dyes switching.
- looked at imaged F7-647,F6-750 samples, also detect no staining in 750. Previously observed great 750 spots on this very sample imaged on the 16th and 17th.
- however do detect decent staining still on the F6-750,F7-647 sample not imaged before, even in BME.
- hypothesis: the dyes pop off.
STORM
- imaging F07 (En +2 TAD)