A cooling chain for studies of cryofixed biological specimens by scanning transmission electron microscopy and X-ray microanalysis.
A cooling chain is described which enables the transfer of frozen hydrated biological specimens (ultrathin cryosections as well as about 1 micrometer thick cultured cells) from a cryoultramicrotome into a scanning transmission electron microscope with a field emission gun. Transfer is done at 118 K, specimen temperature in the microscope is 165 K. Sublimation processes are controlled visually and by mass spectrometry. Electron micrographs and X-ray microanalytical spectra of cryofixed unstained tissue culture cells and rat liver tissue sections are described and discussed. Contamination of the specimen is much reduced by use of the cold stage.
D000818:Animals; D002478:Cells, Cultured / Q000648:ultrastructure; D003584:Cytological Techniques; D004577:Electron Probe Microanalysis / Q000295:instrumentation*; D005615:Freezing; D008099:Liver / Q000648:ultrastructure; D008855:Microscopy, Electron, Scanning / Q000295:instrumentation*; D008867:Microtomy / Q000379:methods*; D051381:Rats