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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.

Original publication

DOI

10.1073/pnas.0400088101

Type

Journal article

Journal

Proc Natl Acad Sci U S A

Publication Date

09/03/2004

Volume

101

Pages

3346 - 3351

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