Orientating the Future Bio-Macromolecular Electron Microscopy

Air Water Interface ◽

The Future

With forty years of developments, bio-macromolecule cryo-electron microscopy has met its revolution of resolution and is playing a very important role in structural biology study. According to different specimen states, cryo-electron microscopy (cryo-EM) involves three specific techniques, single particle analysis (SPA), electron tomography and sub-tomogram averaging, and electron diffraction. All these three techniques have not realized their full potentials of solving structures of bio-macromolecules and therefore need to be developed in the future. In this review, the current existing bottlenecks of cryo-EM SPA are discussed with theoretical analysis, which includes air-water interface during specimen cryo-vitrification, bio-macromolecular conformational heterogeneity, focus gradient within thick specimen, and electron radiation damage. Besides, potential solutions of these bottlenecks are proposed and discussed, which are worthy of further investigations in the future.

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

Science ◽

10.1126/science.aat4346 ◽

2018 ◽

Vol 361 (6405) ◽

pp. 876-880 ◽

Cited By ~ 105

Author(s):

Yifan Cheng

Keyword(s):

Electron Microscopy ◽

Structural Biology ◽

Single Particle ◽

Electron Crystallography ◽

The Past ◽

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.

Recent Developments of Single Particle Analysis with Cryo-electron Microscopy

Nihon Kessho Gakkaishi ◽

10.5940/jcrsj.60.230 ◽

2018 ◽

Vol 60 (5-6) ◽

pp. 230-232

Author(s):

Takanori NAKANE

Keyword(s):

Electron Microscopy ◽

Single Particle ◽

Particle Analysis ◽

Recent Developments

1PT180 Single particle analysis of 26S proteasome by cryo-electron microscopy(The 50th Annual Meeting of the Biophysical Society of Japan)

Seibutsu Butsuri ◽

10.2142/biophys.52.s99_5 ◽

2012 ◽

Vol 52 (supplement) ◽

pp. S99-S100

Author(s):

Kaoru Mitsuoka ◽

Daisuke Kasuya ◽

Yasushi Saeki ◽

Takuo Yasunaga

Keyword(s):

Electron Microscopy ◽

Annual Meeting ◽

Single Particle ◽

26S Proteasome ◽

Particle Analysis ◽

Biophysical Society

Using Cryo-EM to Investigate Bacterial Secretion Systems

Annual Review of Microbiology ◽

10.1146/annurev-micro-090817-062702 ◽

2018 ◽

Vol 72 (1) ◽

pp. 231-254 ◽

Cited By ~ 3

Author(s):

Chiara Rapisarda ◽

Matteo Tassinari ◽

Francesca Gubellini ◽

Rémi Fronzes

Keyword(s):

Electron Microscopy ◽

Particle Analysis ◽

Secretion Systems ◽

Extracellular Milieu ◽

Bacterial Fitness ◽

High Resolution Images ◽

The Impact ◽

Bacterial Secretion

Bacterial secretion systems are responsible for releasing macromolecules to the extracellular milieu or directly into other cells. These membrane complexes are associated with pathogenicity and bacterial fitness. Understanding of these large assemblies has exponentially increased in the last few years thanks to electron microscopy. In fact, a revolution in this field has led to breakthroughs in characterizing the structures of secretion systems and other macromolecular machineries so as to obtain high-resolution images of complexes that could not be crystallized. In this review, we give a brief overview of structural advancements in the understanding of secretion systems, focusing in particular on cryo–electron microscopy, whether tomography or single-particle analysis. We describe how such techniques have contributed to knowledge of the mechanism of macromolecule secretion in bacteria and the impact they will have in the future.