Difference between revisions of "With FACS"

From Tanaka Wiki
Jump to navigation Jump to search
 
Line 11: Line 11:
 
*Count the cells
 
*Count the cells
  
**The sample is ready for downstream applications:
+
*The sample is ready for downstream applications:
***[[For_scRNA-seq|scRNA-seq]]
+
**[[For_scRNA-seq|scRNA-seq]]
***[[For_transplantation|cell transplantation]]
+
**[[For_transplantation|cell transplantation]]

Latest revision as of 21:10, 18 July 2022

For FACS


  • Finish the dissociation protocol
  • Resuspend in 150-200uL AMEM(0)
  • Send to FACS facility immediately
    • Book FACS machine F02 because it has both GFP and mCherry lasers.
    • Normal setting: 85um nozzle with 0.7x FACS Flow for frog cells; 100um nozzle with 0.7x FACS Flow for axolotl cells.
  • Collect the cells into a 1.5mL tube with 700uL AMEM(0) inside. Note down the FACS count.
  • Spin down the cells. 300xrcf, 20 degree, 5 minutes.
  • Resuspend in 50-100uL AMEM(0)
  • Count the cells