Blotting-free and lossless cryo-electron microscopy grid preparation from nanoliter-sized protein samples and single-cell extracts

doi:10.1016/j.jsb.2016.11.002

. 2017 Mar;197(3):220-226.

doi: 10.1016/j.jsb.2016.11.002. Epub 2016 Nov 15.

Blotting-free and lossless cryo-electron microscopy grid preparation from nanoliter-sized protein samples and single-cell extracts

Stefan A Arnold¹, Stefan Albiez², Andrej Bieri², Anastasia Syntychaki², Ricardo Adaixo², Robert A McLeod², Kenneth N Goldie², Henning Stahlberg², Thomas Braun³

Affiliations

¹ Center for Cellular Imaging and NanoAnalytics (C-CINA), Biozentrum, University of Basel, Switzerland; Swiss Nanoscience Institute, University of Basel, Switzerland.
² Center for Cellular Imaging and NanoAnalytics (C-CINA), Biozentrum, University of Basel, Switzerland.
³ Center for Cellular Imaging and NanoAnalytics (C-CINA), Biozentrum, University of Basel, Switzerland. Electronic address: thomas.braun@unibas.ch.

PMID: 27864160
DOI: 10.1016/j.jsb.2016.11.002

Free article

Blotting-free and lossless cryo-electron microscopy grid preparation from nanoliter-sized protein samples and single-cell extracts

Stefan A Arnold et al. J Struct Biol. 2017 Mar.

Free article

. 2017 Mar;197(3):220-226.

doi: 10.1016/j.jsb.2016.11.002. Epub 2016 Nov 15.

Authors

Stefan A Arnold¹, Stefan Albiez², Andrej Bieri², Anastasia Syntychaki², Ricardo Adaixo², Robert A McLeod², Kenneth N Goldie², Henning Stahlberg², Thomas Braun³

Affiliations

¹ Center for Cellular Imaging and NanoAnalytics (C-CINA), Biozentrum, University of Basel, Switzerland; Swiss Nanoscience Institute, University of Basel, Switzerland.
² Center for Cellular Imaging and NanoAnalytics (C-CINA), Biozentrum, University of Basel, Switzerland.
³ Center for Cellular Imaging and NanoAnalytics (C-CINA), Biozentrum, University of Basel, Switzerland. Electronic address: thomas.braun@unibas.ch.

PMID: 27864160
DOI: 10.1016/j.jsb.2016.11.002

Abstract

We present a sample preparation method for cryo-electron microscopy (cryo-EM) that requires only 3-20nL of sample to prepare a cryo-EM grid, depending on the protocol used. The sample is applied and spread on the grid by a microcapillary. The procedure does not involve any blotting steps, and real-time monitoring allows the water film thickness to be assessed and decreased to an optimum value prior to vitrification. We demonstrate that the method is suitable for high-resolution cryo-EM and will enable alternative electron microscopy approaches, such as single-cell visual proteomics.

Keywords: Cryo electron microscopy; Microfluidics; Sample preparation.

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Blotting-free and lossless cryo-electron microscopy grid preparation from nanoliter-sized protein samples and single-cell extracts

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Blotting-free and lossless cryo-electron microscopy grid preparation from nanoliter-sized protein samples and single-cell extracts

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