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Self-pressurized rapid rreezing as cryo-fixation method for electron microscopy and cryopreservation of living cells

Reduction or complete prevention of ice crystal formation during freezing of biological 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 micros...

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Hauptverfasser: Hübinger, Jan (VerfasserIn) , Grabenbauer, Markus (VerfasserIn)
Dokumenttyp: Article (Journal)
Sprache:Englisch
Veröffentlicht: 11 May 2018
In: Current protocols in cell biology
Year: 2018, Jahrgang: 79, Heft: 1
ISSN:1934-2616
DOI:10.1002/cpcb.47
Online-Zugang:Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1002/cpcb.47
Verlag, lizenzpflichtig, Volltext: https://currentprotocols.onlinelibrary.wiley.com/doi/abs/10.1002/cpcb.47
Volltext
Verfasserangaben:Jan Huebinger and Markus Grabenbauer

MARC

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520 |a Reduction or complete prevention of ice crystal formation during freezing of biological 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 and cryopreservation is described. Samples are rapidly cooled in tightly sealed metal tubes of high thermal diffusivity and then plunged into a liquid cryogen. Due to the fast cooling speed and high-pressure buildup internally in the confined volume of the metal tubes, ice crystal formation is reduced or completely prevented, resulting in vitrification of the sample. For cryopreservation, however, a similar principle applies to prevent ice crystal formation during re-warming. A detailed description of procedures for cooling (and re-warming) of biological samples using this technique is provided. © 2018 by John Wiley & Sons, Inc. 
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