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Self-Blotting Nanowire Grids for Cryo-EM Sample Preparation

Published online by Cambridge University Press:  04 August 2017

Hui Wei
Affiliation:
National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, USA
Venkat Dandey
Affiliation:
National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, USA
Zhening Zhang
Affiliation:
National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, USA
Ashleigh Raczkowski
Affiliation:
National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, USA
Bridget Carragher
Affiliation:
National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, USA
Clinton S. Potter
Affiliation:
National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, USA

Abstract

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Almost every aspect of cryo electron microscopy (CryoEM) has been automated over the last few decades. One of the challenges that remains to be addressed is the robust and reliable preparation of vitrified specimens of suitable ice thickness. The development of a new self-blotting nanowire (Zhang et al., 2013) grid in conjunction with a piezo electric dispensing robot called Spotiton (Jain et al., 2012) enables spreading a sample to a thin film without the use of externally applied filter paper. This new approach has the advantage of using small amounts of protein material, resulting in large areas of ice of a well- defined thickness containing evenly distributed particles (Razinkov et al., 2016).

Type
Abstract
Copyright
© Microscopy Society of America 2017 

References

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