Biological Applications at the Cutting Edge of Cryo-Electron Microscopy

• Type of Material: Article
• Authors: Rebecca S. Dillard, Emory University; Cheri M. Hampton, Emory University; Joshua D. Strauss, Emory University; Zunlong Ke, Emory University; Deanna Altomara, Emory University; Ricardo C. Guerrero-Ferreira, Emory University; Gabriella Kiss, Emory University; Elizabeth R. Wright, Emory University
• Language: English
• Date: 2018-08-01
• Publisher: Cambridge University Press (CUP): STM Journals
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © Microscopy Society of America 2018
• Final Published Version (URL): http://dx.doi.org/10.1017/S1431927618012382
• Title of Journal or Parent Work: Microscopy and Microanalysis
• ISSN: 1431-9276
• Volume: 24
• Issue: 4
• Start Page: 406
• End Page: 419
• Grant/Funding Information: Public health service grant F32GM112517 to J.D.S. from the NIH.; Reovirus data collection at Florida State University was made possible by NIH grants S10 OD018142-01, S10 RR025080-01, and U24 GM116788 to K. A. T.; This work was supported in part by Emory University, Children’s Healthcare of Atlanta, and the Georgia Research Alliance to E.R.W.; the Center for AIDS Research at Emory University (P30 AI050409); the James B. Pendleton Charitable Trust to E.R.W.; public health service grants R01GM104540, R01GM114561, R21AI101775, R01GM104540-03S1 to E.R.W. from the NIH, and NSF grant 0923395 to E.R.W.
• Keywords: SAMPLE PREPARATION; cryo-correlative light and electron microscopy (cryo-CLEM); CRYO-EM STRUCTURE; Microscopy; MOLECULAR ARCHITECTURE; RESOLUTION ELECTRON CRYOMICROSCOPY; FOCUS PHASE-CONTRAST; RAPID FREEZING SPRF; vitrification; cryo-electron tomography (cryo-ET); BEAM-INDUCED MOTION; CORRELATIVE LIGHT; phase plates; direct electron detectors; FLUORESCENCE LIGHT-MICROSCOPY; Science & Technology; Materials Science, Multidisciplinary; Technology; Materials Science; cryo-electron microscopy (cryo-EM); ANGSTROM STRUCTURE; transmission electron microscopy (TEM)
• Research Categories: Biology, Microbiology; Health Sciences, Immunology

Abstract

Cryo-electron microscopy (cryo-EM) is a powerful tool for macromolecular to near-atomic resolution structure determination in the biological sciences. The specimen is maintained in a near-native environment within a thin film of vitreous ice and imaged in a transmission electron microscope. The images can then be processed by a number of computational methods to produce three-dimensional information. Recent advances in sample preparation, imaging, and data processing have led to tremendous growth in the field of cryo-EM by providing higher resolution structures and the ability to investigate macromolecules within the context of the cell. Here, we review developments in sample preparation methods and substrates, detectors, phase plates, and cryo-correlative light and electron microscopy that have contributed to this expansion. We also have included specific biological applications.