DOI: 10.1063/1.4967864. A precision cryostat design for manual and semi-automated cryo-plunge instruments

A precision cryostat design for manual and semi-automated cryo-plunge instruments

10.1063/1.4967864

Crossref journal-article

AIP Publishing

Review of Scientific Instruments (317)

Abstract

Here we describe a bench-top cryostat system to control the temperature of liquid ethane in a cryo-plunge apparatus designed for biological specimen preparation for electron cryomicroscopy. It comprises a foam insulated Dewar containing a copper cryostat cup, whose temperature is controlled via an active feedback system to within 0.1 K. The device can easily be incorporated into existing manual and semi-automatic cryo-plunge instruments that are not equipped with cryogenic temperature control. Over the course of normal use, we find that using a cryostat is convenient, fast, and does not require special mixtures of cryogens like ethane/propane. This simple cryostat improves the reliability and reproducibility of biological specimen preparation for electron cryomicroscopy.

Bibliography

Russo, C. J., Scotcher, S., & Kyte, M. (2016). A precision cryostat design for manual and semi-automated cryo-plunge instruments. Review of Scientific Instruments, 87(11).

Authors 3

Christopher J. Russo (first)
Steve Scotcher (additional)
Martin Kyte (additional)

References 20 Referenced 48

10.1017/S0033583500004297 / Q. Rev. Biophys. / Cryo-electron microscopy of vitrified specimens (1988)
10.1126/science.1251652 / Science / The resolution revolution (2014)
10.1107/S2052252515023738 / IUCrJ / CryoEM at IUCrJ: A new era (2016)
10.1038/308032a0 / Nature / Cryo-electron microscopy of viruses (1984)
{'key': '2023070419145206300_c5', 'volume-title': 'Handbook of Chemistry and Physics', 'author': 'Haynes', 'year': '2015', 'edition': '96th ed.'} / Handbook of Chemistry and Physics by Haynes (2015)
10.1111/j.1365-2818.1981.tb01224.x / J. Microsc. / The design and use of a simple device for rapid quench-freezing of biological samples (1981)
10.1002/jemt.1060100111 / J. Electron Microsc. Tech. / Controlled environment vitrification system: An improved sample preparation technique (1988)
10.1016/0968-4328(93)90009-P / Micron / Fast-freezing devices for cryo-electron-microscopy (1993)
10.1016/0304-3991(94)90012-4 / Ultramicroscopy / Analysis of transient structures by cryo-microscopy combined with rapid mixing of spray droplets (1994)
10.1016/S0076-6879(05)91024-0 / Methods Enzymol. / Cryoelectron microscopy of liposomes (2005)
10.1016/S0076-6879(10)81003-1 / Methods Enzymol. / Plunge freezing for electron cryomicroscopy (2010)
10.1002/jemt.1060130305 / J. Electron Microsc. Tech. / High-pressure freezing for the preservation of biological structure: Theory and practice (1989)
10.1093/jmicro/dfr044 / Microscopy / The origins and evolution of freeze-etch electron microscopy (2011)
{'volume-title': 'The Montreal Protocol on Substances that Deplete the Ozone Layer', 'year': '1989', 'author': 'The Ozone Secretariat', 'key': '2023070419145206300_c14'} / The Montreal Protocol on Substances that Deplete the Ozone Layer by The Ozone Secretariat (1989)
10.1017/S1431927608080781 / Microsc. Microanal. / An improved cryogen for plunge freezing (2008)
10.1016/j.jsb.2003.09.027 / J. Struct. Biol. / A second generation apparatus for time-resolved electron cryo-microscopy using stepper motors and electrospray (2003)
10.1016/j.jsb.2008.11.007 / J. Struct. Biol. / Implementation of a flash-photolysis system for time-resolved cryo-electron microscopy (2009)
10.1016/j.sbi.2015.12.007 / Curr. Opin. Struct. Biol. / Progress towards an optimal specimen support for electron cryomicroscopy (2016)
10.1016/bs.mie.2016.04.011 / Methods Enzymol. / Specimen preparation for high-resolution cryo-em (2016)
10.1126/science.1259530 / Science / Ultrastable gold substrates for electron cryomicroscopy (2014)

Dates

Type	When
Created	8 years, 9 months ago (Nov. 21, 2016, 1 p.m.)
Deposited	2 months, 4 weeks ago (May 23, 2025, 9:09 a.m.)
Indexed	3 weeks, 2 days ago (July 30, 2025, 7:07 a.m.)
Issued	8 years, 9 months ago (Nov. 1, 2016)
Published	8 years, 9 months ago (Nov. 1, 2016)
Published Online	8 years, 9 months ago (Nov. 21, 2016)
Published Print	8 years, 9 months ago (Nov. 1, 2016)

Funders 2

Medical Research Council 10.13039/501100000265
Region: Europe
gov (National government)
Labels3
1. Medical Research Council (United Kingdom)
2. UK Medical Research Council
3. MRC
Awards1
1. U105192715
Leverhulme Trust 10.13039/501100000275
Awards1
1. Early Career Fellowship

BibTeX

@article{Russo_2016, title={A precision cryostat design for manual and semi-automated cryo-plunge instruments}, volume={87}, ISSN={1089-7623}, url={http://dx.doi.org/10.1063/1.4967864}, DOI={10.1063/1.4967864}, number={11}, journal={Review of Scientific Instruments}, publisher={AIP Publishing}, author={Russo, Christopher J. and Scotcher, Steve and Kyte, Martin}, year={2016}, month=nov }

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