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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| Main Authors: | , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
11 May 2018
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| In: |
Current protocols in cell biology
Year: 2018, Volume: 79, Issue: 1 |
| ISSN: | 1934-2616 |
| DOI: | 10.1002/cpcb.47 |
| Online Access: | Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1002/cpcb.47 Verlag, lizenzpflichtig, Volltext: https://currentprotocols.onlinelibrary.wiley.com/doi/abs/10.1002/cpcb.47 
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| Author Notes: | Jan Huebinger and Markus Grabenbauer |
| Summary: | 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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| Item Description: | Gesehen am 30.04.2020 |
| Physical Description: | Online Resource |
| ISSN: | 1934-2616 |
| DOI: | 10.1002/cpcb.47 |