Matt:LabNotes/2015-9-2
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Imaging BA8 with DARTFISH suppv2 for 3D Decoding[edit]
Sample[edit]
- I think this sample has suppv2 because in ConfocalStore Hosuk imaged sample on 5-18-2015 and named it "S2_V4_Supp2_2015-05-18"
- Notes on lid of culture dish:
- 2 5/16/15
- BA8 (4/29/15)
- Fix: 4% Form/37C/15min
- 0.25% TX100/10min/RT
- 0.01%Pepsin/5min/37C
- RT:+adUTP 5/16/15
- 15min@RT->37C
- RCA: 5/18/15
Adding Fiducial Beads[edit]
- Sonicate 1:2000 diluted fluorescent spheres in bath sonicator
- Wash sample with 10nM HEPES
- Add beads to sample and incubate 5min at RT
- Wash sample with 10nM HEPES twice
- Add BS(PEG)9 for 1hr at RT
- Wash with 1X PBS twice
- Add 1X Tris pH 8.0 for 30min at RT
- Wash with 1X PBS twice
Imaging on 9-2-2015[edit]
- Cycle00: Fiducial Beads + FISGA_Adpt_Cy3
- Cycle01-Cycle05
- Leica software crashed before Cycle05 due to trying to save Cycle04 when there was no more disk space left
- Had to re-find positions
- Leica software crashed before Cycle05 due to trying to save Cycle04 when there was no more disk space left
- 4 positions
- 0.3um z step size
- 48 zSlices