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

Methods ◽  
2016 ◽  
Vol 100 ◽  
pp. 3-15 ◽  
Author(s):  
Rebecca F. Thompson ◽  
Matt Walker ◽  
C. Alistair Siebert ◽  
Stephen P. Muench ◽  
Neil A. Ranson
Keyword(s):  
Electron Microscopy ◽  
Sample Preparation ◽  
Structural Biology ◽  
Cryo Electron Microscopy

scholarly journals An open-source cryo-storage solution

2017 ◽  
Author(s):  
Eveline Ultee ◽  
Fred Schenkel ◽  
Wen Yang ◽  
Susanne Brenzinger ◽  
Jamie S. Depelteau ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Sample Preparation ◽  
Structural Biology ◽  
Storage Capacity ◽  
Storage System ◽  
Cost Effective ◽  
Wait Times ◽  
Sample Storage ◽  
Cryo Electron Microscopy ◽  
And Storage

AbstractThe field of cryo-electron microscopy is a rapidly growing method in structural biology. With this development, access to cryo-EM facilities becomes a bottleneck that results in long wait times between sample preparation and data acquisition. To improve sample storage, we developed a cryo-storage system with a more efficient and larger storage capacity that enables cryo-sample storage in a highly organized manner. This system is simple to use, cost-effective and easily adaptable for any type of grid box and storage dewar and any size cryo-EM laboratory.

Cryo-EM Is a Powerful Tool, But Helical Applications Can Have Pitfalls

Soft Matter ◽  
2021 ◽  
Author(s):  
Edward Egelman ◽  
Fengbin Wang
Keyword(s):  
Electron Microscopy ◽  
Structural Biology ◽  
Macromolecular Complexes ◽  
Atomic Structures ◽  
Cryo Electron Microscopy ◽  
Main Technique

In structural biology, cryo-electron microscopy (cryo-EM) has emerged as the main technique for determining the atomic structures of macromolecular complexes. This has largely been due to the introduction of direct...

scholarly journals Single-particle cryo-EM—How did it get here and where will it go

Science ◽  
2018 ◽  
Vol 361 (6405) ◽  
pp. 876-880 ◽  
Author(s):  
Yifan Cheng
Keyword(s):  
Electron Microscopy ◽  
Structural Biology ◽  
Single Particle ◽  
Electron Crystallography ◽  
The Past ◽  
Cryo Electron Microscopy ◽  
Technological Advances ◽  
The Future ◽  
Electron Cryotomography

Cryo–electron microscopy, or simply cryo-EM, refers mainly to three very different yet closely related techniques: electron crystallography, single-particle cryo-EM, and electron cryotomography. In the past few years, single-particle cryo-EM in particular has triggered a revolution in structural biology and has become a newly dominant discipline. This Review examines the fascinating story of its start and evolution over the past 40-plus years, delves into how and why the recent technological advances have been so groundbreaking, and briefly considers where the technique may be headed in the future.

scholarly journals Rapid high-resolution structure analysis of small, biotechnologically relevant enzymes by cryo-electron microscopy

2021 ◽  
Author(s):  
Nicole Dimos ◽  
Carl P.O. Helmer ◽  
Andrea M. Chanique ◽  
Markus C. Wahl ◽  
Robert Kourist ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
High Resolution ◽  
Structural Biology ◽  
Structure Analysis ◽  
Cryo Electron Microscopy ◽  
High Resolution Structure ◽  
Fast Development ◽  
Pharmaceutical Industries ◽  
Exquisite Specificity ◽  
Resolution Structure

Enzyme catalysis has emerged as a key technology for developing efficient, sustainable processes in the chemical, biotechnological and pharmaceutical industries. Plants provide large and diverse pools of biosynthetic enzymes that facilitate complex reactions, such as the formation of intricate terpene carbon skeletons, with exquisite specificity. High-resolution structural analysis of these enzymes is crucial to understand their mechanisms and modulate their properties by targeted engineering. Although cryo-electron microscopy (cryo-EM) has revolutionized structural biology, its applicability to high-resolution structure analysis of comparatively small enzymes is so far largely unexplored. Here, we show that cryo-EM can reveal the structures of ~120 kDa plant borneol dehydrogenases at or below 2 Å resolution, paving the way for the fast development of new biocatalysts that provide access to bioactive terpenes and terpenoids.

Tailoring surface acoustic wave atomisation for cryo-electron microscopy sample preparation

Lab on a Chip ◽  
2019 ◽  
Vol 19 (8) ◽  
pp. 1378-1385 ◽  
Author(s):  
Dariush Ashtiani ◽  
Alex de Marco ◽  
Adrian Neild
Keyword(s):  
Electron Microscopy ◽  
Fluid Flow ◽  
Sample Preparation ◽  
Acoustic Wave ◽  
Surface Acoustic Wave ◽  
Flow Rate ◽  
Fluid Flow Rate ◽  
Cryo Electron Microscopy ◽  
Electron Microscopy Grid

Surface acoustic wave (SAW) atomisation is investigated in the context of cryo electron microscopy grid preparation. Here, the primary requirements are a reproducible and narrow plume of droplets delivering a low fluid flow rate.

Uncovering the structures of modular polyketide synthases

2015 ◽  
Vol 32 (3) ◽  
pp. 436-453 ◽  
Author(s):  
Kira J. Weissman
Keyword(s):  
Electron Microscopy ◽  
Structural Biology ◽  
Structural Characterization ◽  
Small Angle ◽  
Single Particle ◽  
Polyketide Synthases ◽  
X Ray ◽  
X Ray Scattering ◽  
Cryo Electron Microscopy

This review covers a breakthrough in the structural biology of the gigantic modular polyketide synthases (PKS): the structural characterization of intact modules by single-particle cryo-electron microscopy and small-angle X-ray scattering.

scholarly journals Antibody-Based Affinity Cryo-Electron Microscopy at 2.6 Å Resolution

2016 ◽  
Author(s):  
Guimei Yu ◽  
Kunpeng Li ◽  
Pengwei Huang ◽  
Xi Jiang ◽  
Wen Jiang
Keyword(s):  
Electron Microscopy ◽  
Sample Preparation ◽  
High Concentration ◽  
Low Concentration ◽  
Atomic Structures ◽  
Cryo Electron Microscopy ◽  
Grid Approach ◽  
Reconstruction Quality ◽  
Tulane Virus ◽  
Quantitative Analyses

AbstractThe affinity cryo-electron microscopy (cryo-EM) approach has been explored in recent years to simplify and improve the sample preparation for cryo-EM. Despite the demonstrated successes for low-concentration and unpurified specimens, the lack of near-atomic structures using this approach has led to a common perception of affinity cryo-EM as a niche technique incapable of reaching high resolutions. Here, we report a ~2.6 Å structure solved using the antibody-based affinity grid approach with a Tulane virus sample of low concentration. This is the first near-atomic structure solved using the affinity cryo-EM approach. Quantitative analyses of the structure indicate data and reconstruction quality comparable to conventional grid preparation method using samples at high concentration. With the shifting of bottlenecks of cryo-EM structural studies to sample grid preparation, our demonstration of the sub-3 Å capability of affinity cryo-EM approach indicates its potential in revolutionizing cryo-EM sample preparation for a broader spectrum of specimens.

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