9:00a – 5:00p
- Guest speaker, see post.
Multicolor STORM more trouble-shooting
- observed that 647 images look particularly weak, especially given the brightness of these spots.
- 15 min of max 405 completely bleaches the 647 spot. (~50K frames).
- 561 laser at 10% is .45-5 mW at back objective at 35 mW power source.
- 405 laser at max power is .45 mW at back objective
- 40 min of 461 at .45 mW (10% AOTF, 35mW source set), results in about a 2/3rds reduction in brightness of the spots.
- should test 488.
- imaging depth — 750 now appears above instead of below the 647 channel? (at least in test image, 750 spot visible in same x-y position when offset is 200 above the 647 position). This is very weird, the beads don’t seem to have this offset. maybe a
- area bleaching is stronger / larger destroyed region in 2 color samples. space out the fields of view more.
- 647 greatly-greatly diminished after 750 imaging. Long time at max 405 power appears to be the cause.
- can’t excite simultaneously — too much blead-through.
- just keep the 405 low and image for longer.
- see protected post
- Start processing of data on Cajal and Tuck for L3F07, L3F08, L3E04, L3E04E05, and L3E06E07
- resumed work on integrating new ChromeWarp method into STORMfinder