Scanning electron microscopy of cells and tissues under fully hydrated conditions.
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.
Thiberge
Stephan
S
Department of Physics of Complex Systems, The Weizmann Institute of Science, Rehovot 76100, Israel.
Nechushtan
Amotz
A
Sprinzak
David
D
Gileadi
Opher
O
Behar
Vered
V
Zik
Ory
O
Chowers
Yehuda
Y
Michaeli
Shulamit
S
Schlessinger
Joseph
J
Moses
Elisha
E
D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003584:Cytological Techniques / Q000379:methods*; D004583:Electrons; D006046:Gold; D006367:HeLa Cells; D006652:Histological Techniques / Q000379:methods*; D006801:Humans; D051379:Mice; D008855:Microscopy, Electron, Scanning / Q000379:methods*; D012542:Scattering, Radiation; D013194:Staining and Labeling / Q000379:methods; D014346:Trypanosoma brucei brucei / Q000648:ultrastructure; D014867:Water
D014867:Water; D006046:Gold