Advertisement

Scanning Electron Microscopy

  • Michael J. Dykstra
  • Laura E. Reuss

Abstract

Scanning electron microscopy (SEM) has a history almost as old as TEM, but the development of a commercial product took much longer. (1938) built the first SEM, and (1942) produced an SEM with a 50-nm probe. A group in Cambridge, England headed by Oatley began work in 1948 that led to the first commercial SEM (the Cambridge Stereoscan) in 1965. (1963) achieved a beam diameter of 5nm, which resulted in 10-nm resolution. With the introduction of the Cambridge Stereoscan in 1965, the biological Community immediately began exploiting the tool to examine numerous tissues and cells. During the 1970s, resolution was improved to 5–6 nm, microprobe analysis (EDS) was applied to SEMs, freeze-fracture methods were explored (Haggis, 1972), and magnifications of over 100,000 × became a reality. The 1980s saw the introduction of FEGs in commercially produced SEMs offered by Hitachi and then JEOL, which allowed lower accelerating voltages and increased magnification and resolution (under 1 nm) and which were immediately adopted by materials scientists. A major technological advance occurred in 1986, when Cambridge Instruments introduced the first digital SEM. This allowed images to be stored and transmitted as digital files, allowed several independently captured images to be averaged to minimize charging artifacts, and led to the contemporary assortment of SEMs operated through a Microsoft Windows™ environment, which has provided a user platform that is accessible to most scientists at work today. By the late 1980s, biologists had discovered FEG instruments and were examining tissues and cells at low voltages with high magnifications previously possible only with sectioned material viewed with a TEM (Ris, 1991). The 1980s and 1990s also saw improvements in probe diameters for microanalysis along with better instrument sensitivity, allowing recognition of elements with lower Z-numbers than previously possible (see Chapter 14).

Keywords

Specimen Surface Backscatter Electron Primary Electron Beam Specimen Chamber Conventional SEMs 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Apkarian, R.P. 1989. Condenser/objective lens SE-I imaging of chromium coated biological specimens using a Schottky field emission source. In: Proc. 47th Ann. Meeting Electron Microsc. Soc. Am. (p. 68). San Francisco Press, San Francisco.Google Scholar
  2. Boyes, E.D. 2000. On low voltage scanning electron microscopy and chemical microanalysis. Microsc. Microanal. 6: 307.Google Scholar
  3. Bray, D.F., Bagu, J., and Koegler, P. 1993. Comparison of hexamethyldisilazane (HMDS), Peldri II and critical-point drying methods for scanning electron microscopy of biological specimens. Microsc. Res. Tech. 26: 489.Google Scholar
  4. Claugher, D. (ed.). 1990. Scanning electron microscopy in taxonomy and functional morphology. Clarendon Press, Oxford.Google Scholar
  5. Crang, R.F.E., and Klomparens, K.L. 1988. Artifacts in biological electron microscopy. Plenum, New York.Google Scholar
  6. Dey, S., Basu Baul, TS., Roy, B., and Dey, D. 1989. A new rapid method of air-drying for scanning electron microscopy using tetramethylsilane. J. Microsc. 156: 259.Google Scholar
  7. Edwards, D.F., Patton, CS., Bemis, D.A., Kennedy, J.R., and Selcer, B.A. 1983. Immotile cilia Syndrome in three dogs from a litter. J. Am. Vet. Med. Assoc. 183: 667.Google Scholar
  8. Goldstein, J.I., Newbury, D.E., Echlin, P., Joy, D.C., Romig, A.D., Jr., Lyman, C.E., Fiori, C., and Lifshin, E. 1992. Scanning electron microscopy and X-ray microanalysis, 2nd edn. Plenum, New York.Google Scholar
  9. Haggis, G.H. 1972. Freeze-fracture for scanning electron microscopy. In: Proc. Fiflh Eur. Congress Electron Microsc. (p. 250). The Institute of Physics, London.Google Scholar
  10. Hainfeld, J.F. 1977. Understanding and using field emission sources. Scan Electron Microsc. 1: 591.Google Scholar
  11. Hanstede, J.G., and Gerrits, P.O. 1982. A new plastic for morphometric investigation of blood vessels, especially in large organs such as the human liver. Anat. Rec. 203: 307.Google Scholar
  12. Hayat, M.A. 1978. Introduction to biological scanning electron microscopy. University Park Press, Baltimore.Google Scholar
  13. Hossler, FE., and Douglas, J.E. 2001. Vascular corrosion casting: Review of advantages and limitations in the application of some simple quantitative methods. Microsc. Microanal. 7: 253.Google Scholar
  14. Humphreys, W.J., Spurlock, B.O., and Johnson, J.S. 1975. Transmission electron microscopy of tissue prepared for scanning electron microscopy by ethanol-cryofracturing. Stain Tech. 50: 119.Google Scholar
  15. Kan, F.W.K. 1990. Use of Peldri II as a Sublimation dehydrant in place of critical-point drying in fracture-label cytochemistry and in backscattered electron imaging fracture-label. J. Electron Microsc. Tech. 14: 21.Google Scholar
  16. Kelley, R.O., Dekker, R.A.F., and Bluemink, J.G. 1975. Thiocarbohydrazide-mediated osmium binding: A technique for protecting soft biological specimens in the scanning electron microscope. In: M.A. Hayat (ed.), Principles and techniques of scanning electron microscopy: Biological applications, Vol. IV. Van Nostrand-Reinhold, New York.Google Scholar
  17. King, E.J., and Brown, M.F. 1983. A technique for preserving aerial fungal structures for scanning electron microscopy. Can. J. Microbiol. 29: 653.Google Scholar
  18. Mazia, D., Schatten, G., and Sale, W. 1975. Adhesion of cells to surfaces coated with polylysine. Applications to electron microscopy. J. Cell Biol. 66: 198.Google Scholar
  19. Motta, P.M., Murakami, T, and Fujita, H. 1992. Scanning electron microscopy of vascular casts: Methods and applications. Kluwer Academic, Boston.Google Scholar
  20. Nation, J.L. 1983. A new method using hexamethyldisilazane for preparation of soft insect tissues for scanning electron microscopy. Stain Tech. 58: 347.Google Scholar
  21. Okada, S., and Schraufnagel, D.E. 2002. Microvasculature of the olfactory organ in the Japanese monkey (Macaca fuscata fuscata). Microsc. Microanal. 8: 159.Google Scholar
  22. Oshel, P. 1994. Hexamethyldisilazane (HMDS) as a Substitute for critical point drying. Microsc. Today 97–4: 19.Google Scholar
  23. Pawley, J.B. 1988. Low voltage scanning electron microscopy. EMSA Bull. 18: 61.Google Scholar
  24. Pease, R.F.W. 1963. High resolution scanning electron microscopy. Ph.D. dissertation, Cambridge University.Google Scholar
  25. Postek, M.T., Howard, K.S., Johnson, A.H., and McMichael, K.L. 1980. Scanning electron microscopy. A student’s handbook. Ladd Research Industries, Burlington, VT.Google Scholar
  26. Ris, H. 1991. The three-dimensional structure of the nuclear pore complex as seen by high voltage electron microscopy and high resolution low voltage scanning electron microscopy. Electron Microsc. Soc. Am. Bull. 21: 54.Google Scholar
  27. Suzuki, F. 1982. Microvasculature of the mouse testis and excurrent duet system. Am. J. Anat. 163: 309.Google Scholar
  28. Suzuki, T, Shibata, M., Tanaka, K., Tsuchida, K., and Toda, T. 1995. A new drying method: Low-vacuum SEM freeze drying and its application to plankton Observation. Bull. Plankt. Soc. Japan 42: 53.Google Scholar
  29. Von Ardenne, M. 1938. Das Elektronene-rastermikroskop, praktische ausfuhrung. Z. Tech. Phys. 19: 407.Google Scholar
  30. Zworykin, V.K., Hillier, J., and Snyder, R.L. 1942. A scanning electron microscope. Am. Soc. Test. Mater. Bull. 117: 15.Google Scholar

Copyright information

© Michael J. Dykstra 2003

Authors and Affiliations

  • Michael J. Dykstra
    • 1
  • Laura E. Reuss
    • 1
  1. 1.North Carolina State UniversityUSA

Personalised recommendations