A portable cryo-plunger for on-site intact cryogenic microscopy sample preparation in natural environments |
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Authors: | Comolli Luis R Duarte Robert Baum Dennis Luef Birgit Downing Kenneth H Larson David M Csencsits Roseann Banfield Jillian F |
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Affiliation: | Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA. lrcomolli@lbl.gov |
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Abstract: | 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. |
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Keywords: | cryo‐electron microscopy cryo‐plunging environmental microbial communities Archaea extremophiles |
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