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Immunogold Labelling for Scanning Electron Microscopy

  • Martin W. Goldberg
  • Jindriska Fiserova
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 657)

Abstract

Scanning electron microscopes are useful biological tools that can be used to image the surface of whole organisms, tissues, cells, cellular components and macromolecules. Processes and structures that exist at surfaces can be imaged in pseudo or real 3D at magnifications of anything from about ×10 to ×1,000,000. Therefore a whole multicellular organism, such as a fly, or a single protein embedded in one of its cell membranes can be visualised. In order to identify that protein at high resolution, or to see and quantify its distribution at lower magnifications, samples can be labelled with antibodies. Any surface that can be exposed can potentially be studied in this way. Presented here is a generic method for immunogold labelling for scanning electron microscopy, using two examples of specimens: isolated nuclear envelopes and the cytoskeleton of mammalian culture cells. Various parameters for sample preparation, fixation, immunogold labelling, drying, metal coating and imaging are discussed so that the best immunogold scanning electron microscopy results can be obtained from different types of specimens.

Key words

Scanning electron microscopy immunogold labelling Xenopus nuclear envelope 

Notes

Acknowledgements

This work was funded by the Biotechnology and Biological Sciences Research Council, UK grant number BB/E015735/1.

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Martin W. Goldberg
    • 1
  • Jindriska Fiserova
    • 1
  1. 1.School of Biological and Biomedical SciencesDurham UniversityDurhamUK

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