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- Title
Imaging of intramembranous particles in frozen-hydrated cells ( Saccharomyces cerevisiae) by high resolution cryo SEM.
- Authors
Walther, P.; Hentschel, J.; Herter, P.; Müller, T.; Zierold, K.
- Abstract
In order to investigate the potential of high-resolution cryoscanning electron microscopy (SEM), frozen-hydrated bulk specimens were compared with transmission electron microscopy (TEM) freeze-fracture replicas. For this purpose a field emission SEM was equipped with a cryo unit consisting of a cold stage in the electron microscopical column and a dedicated cryo preparation chamber. Baker's yeast, Saccharomyces cerevisiae, was the chosen test specimen. The hexagonally arranged intramembranous particles of the plasmic fracture face of the plasmalemma are well known from TEM freeze-fracture studies and can be clearly discerned from structures deriving from preparation artifacts. After freezing, fracturing, sputter-coating with 3 nm of platinum, and vacuum transfer to the cold goniometer stage, the 16.5 nm periodic structure was resolved in the cryo-SEM. Also, intramembranous particles in the nuclear envelope are preserved. Typical artifacts of cryo-SEM are water vapour contaminations and mass loss due to electron irradiation of ice. However, these artifacts are easily recognized and discerned from biological information. Specific advantages of cryo-SEM of frozen-hydrated specimens are: (1) In comparison to SEM of dried material, dehydration artifacts such as shrinkage can be excluded. (2) Compared with freeze-fracture replicas, larger specimens are obtained because no replica cleaning is necessary.
- Publication
Scanning, 1990, Vol 12, Issue 6, p300
- ISSN
0161-0457
- Publication type
Article
- DOI
10.1002/sca.4950120604