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An introduction to sample preparation and imaging by cryo-electron microscopy for structural biology

DOI: 10.1016/j.ymeth.2016.02.017 DOI Help

Authors: Rebecca F. Thompson (University of Leeds) , Matt Walker (MLW Consulting) , C. Alistair Siebert (Electron Bio-Imaging Centre, Diamond Light Source) , Stephen P. Muench (University of Leeds) , Neil A. Ranson (University of Leeds)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Methods

State: Published (Approved)
Published: February 2016

Open Access Open Access

Abstract: Transmission electron microscopy (EM) is a versatile technique that can be used to image biological specimens ranging from intact eukaryotic cells to individual proteins greater than 150 kDa. There are several strategies for preparing samples for imaging by EM, including negative staining and cryogenic freezing. In the last few years, cryo-EM has undergone a ’resolution revolution’, owing to both advances in imaging hardware, image processing software, and improvements in sample preparation, leading to growing number of researchers using cryo-EM as a research tool. However, cryo-EM is still a rapidly growing field, with unique challenges. Here, we summarise considerations for imaging of a range of specimens from macromolecular complexes to cells using EM.

Journal Keywords: eBIC; Electron microscopy; cryo-EM

Subject Areas: Biology and Bio-materials

Diamond Offline Facilities: Electron Bio-Imaging Centre (eBIC)
Instruments: Krios I-Titan Krios I at Diamond

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