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*[[Hosuk:Lab_Notes|LabNote]] ===Rolonies with fresh cells (96well plate)=== ====Process==== *Start ([[Hosuk:Lab_Notes/2013-10-17|10/17]]) *End RCA : 1:30pm 10/19 ====Result==== *16bit 2048 x 2048 images *Not good : there are rolonies, but almost no ACTB --> Max Projection image [[Media:Series032_Stack_Cy5_MaxProjection.tif|1st Rolonies]], [[Media:Series032_Stack_ATTO488_MaxProjection.tif|ACTB]] *Cells in 6 out of 8 wells don’t look good, they were detached, less cells in wells.. WHY? Something happened when fixing? *Cells were aggregated or became chunk… --> need to mix when split *Fixing two step : 37% Formalin --> 10% Formalin *The very first protocol was two step fixing, but Jonathan has been doing one step (10% Formalin), and in their manuscript as well. *But… I want to try two step fixing and compare to one step fixing…
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