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Techniques

  • Michael J. Dykstra
  • Laura E. Reuss

Abstract

Fixation of biological samples for SEM, in general, utilizes the same principles and materials as described for preparing samples for TEM in Chapter 1. Osmium tetroxide postfixation can usually be omitted, though samples that have “charging problems” leading to image distortion can frequently benefit from osmication. Subsequent dehydration in an ethanol or acetone series followed by critical point drying, freeze-drying, or drying with one of the chemical techniques described in this section produces a sample suitable for viewing with a conventional high-vacuum SEM. Preparation of samples for ESEM or LV SEM viewing may not require dehydration, drying, and coating protocols used for high-vacuum SEM, while materials destined for microanalytical techniques may necessitate specialized techniques to avoid spurious signal generation.

Keywords

Vent Valve Sodium Metaperiodate Aldehyde Fixative Free Aldehyde Group Sight Glass 
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.

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Reference

  1. Mazia, D., Schatten, G., and Sale, W. 1975. Adhesion of cells to surfaces coated with polylysine. J. Cell Biol. 66: 198.Google Scholar

Reference

  1. Clayton, D.F. and Alvarez-Buylla, A. 1989. In situ hybridization using PEG-emfoedded tissue and riboprobes: Increased cellular detail coupled with high sensitivity. J. Histochem. Cytochem. 37: 389.Google Scholar

References

  1. 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
  2. Nation, J.L. 1983. A new method using hexamethyldisilazane for preparation of soft insect tissues for scanning electron microscopy. Stain Tech. 58: 347–351.Google Scholar
  3. Oshel, P. 1997. Hexamethyldisilazane (HMDS) as a Substitute for critical point drying. Microsc. Today 97-4: 19.Google Scholar

References

  1. Motta, P.M., Murakami, T, and Fujita, H (eds.). 1992. Scanning electron microscopy of vascular casts: Methods and applications. Kluwer Academic, Boston.Google Scholar
  2. Suzuki, F. 1982. Microvasculature of the mouse testis and excurrent duct system. Am. J. Anat. 163: 309–325.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

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