Sectioning --- frozen tissue

1. Fill the base mold with frozen tissue matrix.

2. Place tissues in the base molds filled with frozen tissue matrix.

3. Place the base mold in 2-methylbutane pre-chilled with liquid nitrogen until the block ALMOST solidifies (30 seconds).

4. Remove tissue block from 2-methylbutane and place blocked tissues on dry ice or store at -70 C.

5. Attach the frozen tissue block on the cryostat chuck. Allow tissue block to equilibrate to the cryostat temperature (-20 C) before cutting sections.

6. Cut sections at 4-6 microns and pick up onto slide.

7. Dry at room temperature.

8. Fix in cold acetone (-20 C) for 2 minutes.

9. Dry fixed slides at room temperature.

10. Store at -70C.