10:00 A – 12:45 A
morning tasks
- break down O/N STORM2 run on B + feducials
- transfer data to ProRAID
Slides for XZ meeting
- explain Ph project
- outlook is good
- should compare to Pc clusters
DNA in situs
- 12 min wash in 2x SSCT at 45C. Then check image quality in Trolox on STORM2/TIRF
- B control worked. Locus is definetely polyploid (many cells have 5 spots. Some spots have small daughter spots near by).
- embryo en-locus staining not detectable on Turnkey, looks like decent spots in TIRF on STORM 2. nuclear background still high at 45C.
- G sample worked!
- post wash all samples 2x 12 min at 60C in 2x SSCT
Embryo imaging
- Unable to find same two fields of embryos (found two new fields)
- Take-home lesson 1: really do need to preimage all sections on the coverglass.
- DAPI stain is still strong after hybe. Interfers somewhat with: activator, restricts power to less than 20% 405. This is not so much a problem with activator dye the low 405 power is pretty effective.
- still high background spot contrast. Spots definetely less bright than with cells. could be a locus copy number effect.
STORM
- STORM of several positions which should be +/- En expressing on en embryo slide.
- O/N imaging of G sample with 540 feducials
- O/N STORM analysis on Cajal of B and Y data with feducials