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

Type of publication Peer-reviewed
Publikationsform Original article (peer-reviewed)
Author Arnold Stefan A., Albiez Stefan, Bieri Andrej, Syntychaki Anastasia, Adaixo Ricardo, McLeod Robert A., Goldie Kenneth N., Stahlberg Henning, Braun Thomas,
Project Fast protein-complex isolation, sample preparation and data processing for high-resolution structural analysis and visual proteomics
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Original article (peer-reviewed)

Journal Journal of Structural Biology
Volume (Issue) 197(3)
Page(s) 220 - 226
Title of proceedings Journal of Structural Biology
DOI 10.1016/j.jsb.2016.11.002

Open Access

Type of Open Access Publisher (Gold Open Access)


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.