Abstract
Scanning electron microscopy (SEM) has been almost universally applied for the surface examination and characterization of both natural and man-made objects. Although an invasive technique, developments in electron microscopy over the years has given the microscopist a much clearer choice in how invasive the technique will be. With the advent of low vacuum SEM in the 1970s (The environmental cold stage, 1970) and environmental SEM in the late 1980s (J Microsc 160(pt. 1):9–19, 1989), it is now possible in some circumstances to examine samples without preparation. However, for the examination of biological tissue and cells it is still advisable to chemically fix, dehydrate, and coat samples for SEM imaging and analysis. This chapter aims to provide an overview of SEM as an imaging tool, and a general introduction to some of the methods applied for the preparation of samples.
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References
LaneWC (1970) The environmental cold stage. Proceedings of the Third Scanning Electron Microscopy Symposium, IIT Research Institute, Chicago, IL, 60616
Danilatos G (1989) Mechanisms of detection and imaging in the ESEM. J Microsc 160(pt. 1):9–19
Von Ardenne M (1938) Das Elektronen-Raster Mikroskop (The scanning electron microscope). Z Tech Phys 19:407–416
Von Ardenne M (1938b) Das Elektronenmikroskop; Theoretische Grundlagen (The scanning electron microscope: theoretical fundamentals). Z Phys 109:553–572. Z Techn Phys 19, 407
Moncrieff DA, Barker PR, Robinson VNE (1979) Electron scattering by gas in the scanning electron microscope. J Phys D Appl Phys 12(4):481–488
Craven JP, Baker FS, Thiel BL, Donald AM (2002) Consequences of positive ions upon imaging in low vacuum SEM. J Microsc 205(1):96–105
Newbury DE et al (1987) Advanced scanning electron microscopy and X-ray microanalysis. Plenum Press, New York
Everhart TE, Thornley RFM (1960) Wide-band detector for micro-microampere low-energy electron currents. J Sci Inst 37:246–248
Nasser Hajbagheeri MA (Ed) (1999) Electron microscopy methods and protocols. Methods in Mol Biol Vol 117. Humana Press
Sabatini DD, Bensch K, Barrnett RJ (1963) Cytochemistry and electron microscopy. The preservation of cellular ultrastructure and enzymatic activity by aldehyde fixation. J Cell Biol 17:19–58
Hayat MA (1982) Fixation for electron microscopy. Academic, New York
Nowell JA, Pawley JB (1980) Preparation of experimental animal tissue for SEM. Scan Electron Microsc Pt 2:1–19
Anderson TF (1951) Techniques for the preservation of three dimensional structure in preparing specimens for the electron microscope. Trans NY Acad Sci, Series 2 131:130–134
Conway K, Kieman JA (1999) Chemical dehydration of specimens with 2,2-dimethoxypropane (DMP) for paraffin processing of animal tissues: practical and economic advantages over dehydration in ethanol. Biotech Histochem 74(1):20–26
Carr JH, Anderson RL, Favero MS (1996) Comparison of chemical dehydration and critical point drying for the stabilization and visualization of aging biofilm present on interior surfaces of PVC distribution pipe. J Appl Microbiol 80(2):225–232
Boyde A (1978) Pros and cons of critical point drying and freeze drying for SEM. SEM/1978/II, SEM Inc., AMF O’Hare, IL, pp 303–314
Boyde A, Macoonachie E (1979) Volume changes during preparation of mouse embryonic tissue for scanning electron microscopy. Scanning 2, 149. Witzstrock Verlag Baden-Baden
Witcomb MJ (1981) The suitability of various adhesives as mounting media for scanning electron microscopy. J Microsc 121:289
Stokes DJ Principles and Practice of Variable Pressure/Environmental SEM (VP-ESEM). Royal Microscopical Society. ISBN 978-0-470-06540-2
Tang X, Joy DC (2005) An experimental model of beam broadening in the variable pressure scanning electron microscope. Scanning 27(6):293–297
Jirak J et al (2010) Scintillation SE detector for VP SEM. J Microsc 239(3):233–238
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Jones, C.G. (2012). Scanning Electron Microscopy: Preparation and Imaging for SEM. In: Bell, L. (eds) Forensic Microscopy for Skeletal Tissues. Methods in Molecular Biology, vol 915. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-977-8_1
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DOI: https://doi.org/10.1007/978-1-61779-977-8_1
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