Scanning electron microscopy of cells and tissues under fully hydrated conditions

Stephan Thiberge, Amotz Nechushtan, David Sprinzak, Opher Gileadi, Vered Behar, Ory Zik, Yehuda Chowers, Shulamit Michaeli, Joseph Schlessinger, Elisha Moses

Research output: Contribution to journalArticlepeer-review

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 ≈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 languageEnglish
Pages (from-to)3346-3351
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume101
Issue number10
DOIs
StatePublished - 9 Mar 2004
Externally publishedYes

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