ICC/IF (Cell Climbing Slices)

ICC/IF (Cell Climbing Slices)

  • Place settled coverslip in culture bottle or perforated plate
  • Take out coverslip after cell growth has reached 60%
  • Wash the coverslip 3X with PBS to remove culture medium
  • Immerse the coverslip (cells face up) into cold acetone or 4% paraformaldehyde or neutral formalin for 10 to 20 min (Close the lid to prevent evaporation)
  • Wash the coverslip 3X with PBS
  • Put the coverslip on filter paper (cells face up)
  • Remove the liquid on the coverslip and allow it to dry for 8-10 hrs
  • To thaw the slice, wash with neutral PBS at room temperature for 10-15 min (The cell climbing slice can be stored in gelatin at -20°C for one week.)

Note: This fixation procedure using paraformaldehyde and formalin fixatives may cause autofluorescence in the green spectrum. In this case, you may try fluorophores in the (i) red range or (ii) infrared range if you have an infrared detection system.