1. Fix sample in formalin for 24 hours.

2. Store sample in DI-H₂O.

3. Exchange DI-H₂O for a 30% sucrose solution (some uses graded solution for a few hours each). I usually leave my samples in sucrose for ~12 hours.

4. Soak sample in HistoPrep to allow for embedding media infiltration (may cause sample shrinkage).

5. Mount sample in HistoPrep and allow it to freeze for at least ~15min in the cryostat.

NOTES:

There really is no set protocol for cryosectioning. Some trial and error may be necessary before an optimal system is found for your samples.