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Advanced Techniques in Biological Electron Microscopy III pp 101–166Cite as

Metal Deposition by High-Energy Sputtering for High Magnification Electron Microscopy

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Abstract

Visualization of fine structures of macromolecular dimensions is an important and growing field of modern biological electron microscopy which was until recently exclusively dominated by transmission electron microscopy. However, only specimen domains of subcellular dimensions can be imaged with the transmission electron microscope (TEM) because of instrumental limitations of tilt angles (±60°) and specimen thickness (< 50–100 nm). Specimens compatible with these limitations are replicas of freeze-etched hydrated cells and cell components and deep-etched or dried preparations of isolated molecules. Recent progress made in the development of scanning electron microscopic imaging methods now makes it possible to use also modern scanning electron microscopes (SEM) for high magnification, secondary electron (SE) imaging. Resolution may be increased to dimensions equal to electron beam probe diameters of <1 nm. This improved SEM resolution is similar to that obtained with TEM replicas. The increased SEM tilt capability (±90°) and the high depth of focus, obtained by adjusting the focus while scanning the image (dynamic focusing), enables the SEM to reveal macromolecular fine structures in dried bulk specimens not only of whole single cells but also of complex tissues.

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  1. K.-R. Peters
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  1. Department of Biological Structures School of Medicine, University of Washington, 98195, Seattle, WA, USA

    James K. Koehler Ph. D.

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Peters, KR. (1986). Metal Deposition by High-Energy Sputtering for High Magnification Electron Microscopy. In: Koehler, J.K. (eds) Advanced Techniques in Biological Electron Microscopy III. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-71135-0_3

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