Transmission electron microscopy in molecular structural biology: A historical survey

• Published in: Archives of biochemistry and biophysics Vol. 581; pp. 3 - 18
• Main Author: Harris, J. Robin
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.09.2015
• Subjects: 2D crystal; Cryo-electron microscopy; Digital image analysis; Electron diffraction; History, 20th Century; History, 21st Century; Metal shadowing; Microscopy, Electron, Transmission - history; Microscopy, Electron, Transmission - methods; Molecular Biology - history; Molecular Biology - methods; Negative staining; Single particle averaging; TEM; Thin sectioning; Vitrification; Negative staining; 2D crystal; Electron diffraction; Digital image analysis; Vitrification; Thin sectioning; Single particle averaging; TEM; Metal shadowing; Cryo-electron microscopy
• ISSN: 0003-9861; 1096-0384
• DOI: 10.1016/j.abb.2014.11.011

•This review article covers the historical development and application of TEM.•Emphasis is placed on molecular structural biology.•Early applications in the 1040s through to the beginning of the present century are covered.•The contribution of leading scientists is given due emphasis. In this personal, historic account of macromolecular transmission electron microscopy (TEM), published data from the 1940s through to recent times is surveyed, within the context of the remarkable progress that has been achieved during this time period. The evolution of present day molecular structural biology is described in relation to the associated biological disciplines. The contribution of numerous electron microscope pioneers to the development of the subject is discussed. The principal techniques for TEM specimen preparation, thin sectioning, metal shadowing, negative staining and plunge-freezing (vitrification) of thin aqueous samples are described, with a selection of published images to emphasise the virtues of each method. The development of digital image analysis and 3D reconstruction is described in detail as applied to electron crystallography and reconstructions from helical structures, 2D membrane crystals as well as single particle 3D reconstruction of icosahedral viruses and macromolecules. The on-going development of new software, algorithms and approaches is highlighted before specific examples of the historical progress of the structural biology of proteins and viruses are presented.