Hosuk:LabNotes/2013-10-19: Difference between revisions
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>Hosuki78 (Created page with "*LabNote ===Rolonies with fresh cells (96well plate)=== ====Process==== *Start (10/17) *End RCA : 1:30pm 10/19 *...") |
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*[[Hosuk:Lab_Notes|LabNote]] | *[[Hosuk:Lab_Notes|LabNote]] | ||
===Rolonies with fresh cells (96well plate)=== | ===Rolonies with fresh cells (96well plate)=== | ||
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*Start ([[Hosuk:Lab_Notes/2013-10-17|10/17]]) | *Start ([[Hosuk:Lab_Notes/2013-10-17|10/17]]) | ||
*End RCA : 1:30pm 10/19 | *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… |
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…