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