CRYOPROTECTANT FOR PROPANE JET FREEZING (Ding et al. 1991)
To make 20 ml of buffer for propane jet cryoprotection:
20 mM MES 0.07808 g 195.2
2 mM CaCl2 0.00588 g 147.02
2 mM KCl 0.00298 g 74.557
0.2 M sucrose 1.3692 342.30
Adjust pH to 5.5 using KOH.
Incubate material in the buffer for 5 to 30 minutes before freezing. Ideally, the tissue should be as small as possible. (See requirements of propane jet freezer.)
Ding, B., Turgeon, R. and Parthasarathy, M. V. (1991) Routine cryofixation of plant tissue by propane jet freezing for freeze substitution. Journal of Electron Microscope Technique 19: 107-117.