Self-Pressurized Rapid Freezing as Cryo-Fixation Method for Electron Microscopy and Cryopreservation of Living Cells.

摘要: Reduction or complete prevention of ice crystal formation during freezing biological specimens is mandatory for two important applications: (1) cryopreservation living cells tissues long-term storage, and (2) cryo-fixation ultrastructural investigations by electron microscopy. Here, a protocol that fast, easy-to-use, suitable both described. Samples are rapidly cooled in tightly sealed metal tubes high thermal diffusivity then plunged into liquid cryogen. Due to the fast cooling speed high-pressure buildup internally confined volume tubes, reduced completely prevented, resulting vitrification sample. For cryopreservation, however, similar principle applies prevent re-warming. A detailed description procedures (and re-warming) samples using this technique provided. © 2018 John Wiley & Sons, Inc.