Fixed Cell Pellet

Embedded in Paraffin

  1. Pellet cells by centrifugation 400 g for 10 minutes at room temperature.
  2. Resuspend cells in 10 ml 10% Buffered Formalin, and fix for two hours at room temperature.
  3. Pellet cells by centrifugation 400g for 10 minutes at room temperature.
  4. Resuspend cells in 300 µl of PBS in a microtube.
  5. Prepare 3% low melting point (LMP) agarose in PBS; weigh before microwaving.
  6. Melt in microwave and replace lost water.
  7. Place melted agarose mixture in water bath at 65°. Allow agarose to equilibrate to 65° (about 30 minutes)
  8. Heat microtube with cells in it to 65° for no more than 10 minutes.
  9. Working in water bath, add 600 µl of agarose mixture to cells, and mix by pipetting up and down.
  10. Immediately transfer microtube to a room temperature microcentrifuge, and spin at 2000 g for five minutes.
  11. Place microtube on ice, remove gelled pellet, and trim of excess agarose.
  12. Wrap pellet in lens paper, place in a paraffin processing cassette, and process to paraffin block.