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

• Verfasst von: Hübinger, Jan [VerfasserIn]
• Verfasst von: Grabenbauer, Markus [VerfasserIn]
• Titel: Self-pressurized rapid rreezing as cryo-fixation method for electron microscopy and cryopreservation of living cells
• Verf.angabe: Jan Huebinger and Markus Grabenbauer
• E-Jahr: 2018
• Jahr: 11 May 2018
• Umfang: 13 S.
• Fussnoten: Gesehen am 30.04.2020
• Titel Quelle: Enthalten in: Current protocols in cell biology
• Ort Quelle: New York, NY [u.a.] : Wiley, 1998
• Jahr Quelle: 2018
• Band/Heft Quelle: 79(2018,1) Artikel-Nummer e47, 13 Seiten
• ISSN Quelle: 1934-2616
• DOI: doi:10.1002/cpcb.47
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: cryo-arrest
• Sach-SW: cryo-fixation
• Sach-SW: cryopreservation
• Sach-SW: electron microscopy
• Sach-SW: high-pressure freezing
• Sach-SW: self-pressurized rapid freezing (SPRF)
• Sach-SW: vitrification
• K10plus-PPN: 1696926955

Abstract

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.