Fixing adherent cells

Submitted by kdorfman on Thu, 05/21/2020 - 17:39

To fix the cells grown on a coverslip

  • Work in a fume hood
  • Gently rinse cells 2X in warm PBS (remove medium, add PBS, repeat)
  • Leave cells in this PBS—until you add the fixative.
  • Remove final PBS rinse, add about 1 mL of fixative to the cells
  • Fix cells in hood for 10 mins.
  • Rinse cells by dunking each coverslip in 3 beakers of PBS-Tween-Azide, 10 X per beaker.
  • Place coverslip back in dish with fresh PBS-Tween-Azide
  • If you are not staining immediately, store cells in PBS-Tween-Azide in fridge.


  • Work in the fume hood
  • Wear gloves!
  • Keep track of which side of the coverslip the cells are on
  • Discard used fixative in the appropriate hazardous waste container in the hood.