Hosuk:LabNotes/2013-10-19: Difference between revisions
Jump to navigation
Jump to search
>Hosuki78 No edit summary |
>Hosuki78 No edit summary |
||
Line 10: | Line 10: | ||
====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… | *Cells were aggregated or became chunk… --> need to mix when split | ||
*Fixing two step : 37% 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…