Abstract
Scanning electron microscopes allow us to reach magnifications of 20–130,000× and resolve compositional and topographical images with intense detail. These images are created by bombarding a sample with electrons in a focused manner to generate a black and white image from the electrons that bounce off of the sample. The electrons are detected using positively charged detectors. Scanning electron microscopy permits three-dimensional imaging of desiccated specimens or wet cells and tissues by using variable pressure chambers. SEM ultrastructural analysis and intracellular imaging supplement light microscopy for molecular profiling of prokaryotes, plants, and mammals. This chapter demonstrates how to prepare and image samples that are (a) desiccated and conductive, (b) desiccated and nonconductive but coated with an electron conductive film using a gold sputter coater, and (c) wet and maintained in a hydrated state using a Deben Coolstage.
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References
Phenom Specifications (2016) NanoScience Instruments. http://www.nanoscience.com/products/sem/phenom/phenom-specs/. Accessed 23 April 2016
Fischer ER, Hansen BT, Nair V, Hoyt FH, Dorward DW (2012) Scanning electron microscopy. Curr Protoc Microbiol Chapter 2:Unit 2B 2. doi:10.1002/9780471729259.mc02b02s25
Harisha S (2007) Biotechnology procedures and experiments handbook. Infinity Science Press LLC, Hingham, MA
Talbot MJ, White RG (2013) Cell surface and cell outline imaging in plant tissues using the backscattered electron detector in a variable pressure scanning electron microscope. Plant Methods 9(1):40. doi:10.1186/1746-4811-9-40
Thiberge S, Nechushtan A, Sprinzak D, Gileadi O, Behar V, Zik O, Chowers Y, Michaeli S, Schlessinger J, Moses E (2004) Scanning electron microscopy of cells and tissues under fully hydrated conditions. Proc Natl Acad Sci U S A 101(10):3346–3351. doi:10.1073/pnas.0400088101
Moore DK (2010) The fabulous Phenom: Supplemental Phenom user’s manual, p 285. http://www.nanoscience.com/files/2313/7954/4001/SEM_SimpUserManual.pdf
Quayle K, Clemens G, Sorribes TG, Kinvig HM, Stevenson PG, Conlan XA, Baker MJ (2016) Elemental and molecular profiling of licit, illicit, and niche tobacco. Forensic Sci Int 266:549–554. doi:10.1016/j.forsciint.2016.07.023
Stokes DJ (2003) Recent advances in electron imaging, image interpretation and applications: environmental scanning electron microscopy. Philos Trans A Math Phys Eng Sci 361(1813):2771–2787. doi:10.1098/rsta.2003.1279
Golding CG, Lamboo LL, Beniac DR, Booth TF (2016) The scanning electron microscope in microbiology and diagnosis of infectious disease. Sci Rep 6:26516. doi:10.1038/srep26516
Jacobs JB, Arai M, Cohen SM, Friedell GH (1976) Early lesions in experimental bladder cancer: scanning electron microscopy of cell surface markers. Cancer Res 36(7 PT 2):2512–2517
Im H, Shao H, Park YI, Peterson VM, Castro CM, Weissleder R, Lee H (2014) Label-free detection and molecular profiling of exosomes with a nano-plasmonic sensor. Nat Biotechnol 32(5):490–495. doi:10.1038/nbt.2886
Wang L, Shalek AK, Lawrence M, Ding R, Gaublomme JT, Pochet N, Stojanov P, Sougnez C, Shukla SA, Stevenson KE, Zhang W, Wong J, Sievers QL, MacDonald BT, Vartanov AR, Goldstein NR, Neuberg D, He X, Lander E, Hacohen N, Regev A, Getz G, Brown JR, Park H, Wu CJ (2014) Somatic mutation as a mechanism of Wnt/beta-catenin pathway activation in CLL. Blood 124(7):1089–1098. doi:10.1182/blood-2014-01-552067
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We thank the Manassas campus of Northern Virginia Community College for the use of equipment and materials.
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Nguyen, J.N.T., Harbison, A.M. (2017). Scanning Electron Microscopy Sample Preparation and Imaging. In: Espina, V. (eds) Molecular Profiling. Methods in Molecular Biology, vol 1606. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6990-6_5
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DOI: https://doi.org/10.1007/978-1-4939-6990-6_5
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