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Matt:LabNotes/2015-9-2
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=Imaging BA8 with DARTFISH suppv2 for 3D Decoding= ==Sample== *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=== *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== *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 *4 positions **0.3um z step size **48 zSlices
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