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Scanning electron microscopy of cells and tissues under fully hydrated conditions.

Authors
Journal
Proceedings of the National Academy of Sciences
0027-8424
Publisher
Proceedings of the National Academy of Sciences
Publication Date
Volume
101
Issue
10
Identifiers
DOI: 10.1073/pnas.0400088101
Keywords
  • Animals
  • Cho Cells
  • Cricetinae
  • Cytological Techniques
  • Electrons
  • Gold
  • Hela Cells
  • Histological Techniques
  • Humans
  • Mice
  • Microscopy
  • Electron
  • Scanning
  • Scattering
  • Radiation
  • Staining And Labeling
  • Trypanosoma Brucei Brucei
  • Water
Disciplines
  • Biology
  • Physics

Abstract

A capability for scanning electron microscopy of wet biological specimens is presented. A membrane that is transparent to electrons protects the fully hydrated sample from the vacuum. The result is a hybrid technique combining the ease of use and ability to see into cells of optical microscopy with the higher resolution of electron microscopy. The resolution of low-contrast materials is approximately 100 nm, whereas in high-contrast materials the resolution can reach 10 nm. Standard immunogold techniques and heavy-metal stains can be applied and viewed in the fluid to improve the contrast. Images present a striking combination of whole-cell morphology with a wealth of internal details. A possibility for direct inspection of tissue slices transpires, imaging only the external layer of cells. Simultaneous imaging with photons excited by the electrons incorporates data on material distribution, indicating a potential for multilabeling and specific scintillating markers.

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