DAPI counterstain adherent cells/sections

Contributed by Martin Fitzpatrick, University of Birmingham, United Kingdom

DAPI counterstain of adherent cells or tissue sections on slides

• Perform all fixation, permeabilization and primary staining of your sample before applying the DAPI counterstain. Fixation or permeabilization is not required to stain with DAPI.
• Wash and equilibrate section in PBS for 5 minutes
• Dilute stock DAPI to 300nM concentration in PBS
• Add 300ul of diluted DAPI per spot to the coverslip. Ensure cells/section is completely covered by the droplet.
• Incubate at room temperature for 1-5minutes
• Place sample in PBS bath, washing 3x for 5 minutes then replacing the PBS.
• Add coverslip with appropriate mounting medium