Setup
- Back-optics aligned
- Focus lock working
- Laser power:
- 647 laser at 200 mW source: 130 mW at power meter
- 405 laser: max power = 1.89 mW. 10% = .09 mW
- z-stage drift without focus lock is ~800 nm per min
- drift about 900 nm down during the time it takes to record a z-calibration movie. Utterly unusable.
STORM imaging
- imaged several sections individually. 120,000 frames without needing 405. May need to discard substantial number of frames due to too high emitter density.
- generated overview mosaic on Steve to pick good cell regions
- focus lock froze during O/N imaging. — Consequently z-focus was lost and many sections are blank. :(.
Breakdown
- repeat z-calibration. Prior to calibration, focus holding pretty steady without lock.