4.5 Article

A portable cryo-plunger for on-site intact cryogenic microscopy sample preparation in natural environments

Journal

MICROSCOPY RESEARCH AND TECHNIQUE
Volume 75, Issue 6, Pages 829-836

Publisher

WILEY
DOI: 10.1002/jemt.22001

Keywords

cryo-electron microscopy; cryo-plunging; environmental microbial communities; Archaea; extremophiles

Funding

  1. Office of Science, Office of Biological and Environmental Research, U.S. Department of Energy [DEAC02-05CH11231]

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We present a modern, light portable device specifically designed for environmental samples for cryogenic transmission-electron microscopy (cryo-TEM) by on-site cryo-plunging. The power of cryo-TEM comes from preparation of artifact-free samples. However, in many studies, the samples must be collected at remote field locations, and the time involved in transporting samples back to the laboratory for cryogenic preservation can lead to severe degradation artifacts. Thus, going back to the basics, we developed a simple mechanical device that is light and easy to transport on foot yet effective. With the system design presented here we are able to obtain cryo-samples of microbes and microbial communities not possible to culture, in their near-intact environmental conditions as well as in routine laboratory work, and in real time. This methodology thus enables us to bring the power of cryo-TEM to microbial ecology. Microsc. Res. Tech. 75:829836, 2012. (C) 2011 Wiley Periodicals, Inc.

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