Hosuk:LabNotes/2013-10-19: Difference between revisions

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====Result====
====Result====
*Not good : there are rolonies, but almost no ACTB
*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 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
*Cells were aggregated or became chunk… --> need to mix when split
*Fixing two step : 37% Formalin 10% Formalin
*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.
*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…
*But… I want to try two step fixing and compare to one step fixing…

Latest revision as of 18:25, 22 October 2013

Rolonies with fresh cells (96well plate)[edit]

Process[edit]

  • Start (10/17)
  • End RCA : 1:30pm 10/19


Result[edit]

  • 16bit 2048 x 2048 images
  • Not good : there are rolonies, but almost no ACTB --> Max Projection image 1st Rolonies, 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…