Cells in suspensionΒΆ

Contributed by Martin Fitzpatrick, University of Birmingham, United Kingdom

DAPI counterstain of cells in suspension, e.g. for FACS analysis

  • Dilute stock DAPI mix to a concentration of 3uM in staining buffer, allowing a total of 1ml per sample.
  • Centrifuge cell suspension at 1200rpm for 5 minutes to pellet. Pour off supernatant.
  • Tap/agitate to dislodge and re-suspend cells in remaining buffer.
  • Add 1ml of diluted DAPI to each sample
  • Incubate for 15 minutes at room temperature
  • Analyze cells as required:

For flow cytometry leave cells in DAPI solution.

For fluorescence microscopy pellet cells by centrifugation, remove supernatant and resuspend in PBS. Add a drop of cells to a slide and cover with coverslip.