Protocol: Chemical fixation

On-chip chemical fixation of cultured cells

Materials

  • Cell medium
  • 1X PBS (also used as mounting medium)
  • [Optional] Blocking buffer: PBS with NH4Cl 50 mM; BSA 0.5%
  • Mineral oil for cell culture
  • Stereomicroscope

Outline_protocol.png

Cell culture on VersaLive

Follow the protocol relative to the cell culture on VersaLive.

On-chip chemical fixation

  1. Empty all ports
  2. PBS washing (1x), 20 μL in all ports
  3. 10 min perfusion of PFA 4% in ports #1 to #5 (leave ports A and B empty)
  4. PBS washing (2x), 20 μL in all ports
  5. 5 min perfusion PBS washing, 20 μL in ports #1 to #5 (leave ports A and B empty)
  6. [Optional] 30 min perfusion blocking buffer, 20 μL in ports #1 to #5 (leave ports A and B empty)
  7. Fill all ports with 20 μL PBS + 2.5 μL mineral oil to prevent evaporation, store at 4˚C