2017Arnold BlottingFree

Citation

Arnold, S. A.; Albiez, S.; Bieri, A.; Syntychaki, A.; Adaixo, R.; McLeod, R. A.; Goldie, K. N.; Stahlberg, H. & Braun, T. Blotting-free and lossless cryo-electron microscopy grid preparation from nanoliter-sized protein samples and single-cell extracts. Journal of structural biology, 2017, 197, 220-226

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.

Links

http://www.sciencedirect.com/science/article/pii/S104784771630243X