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首页> 外文期刊>Current Protocols in Cell Biology >Self-Pressurized Rapid Freezing asCryo-Fixation Method for ElectronMicroscopy and Cryopreservation ofLiving Cells
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Self-Pressurized Rapid Freezing asCryo-Fixation Method for ElectronMicroscopy and Cryopreservation ofLiving Cells

机译：用于电子镜和卵形冷冻保存的自加压快速冻结Ascryo固定方法

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摘要

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