Frozen‐hydrated and Drying Thin Cryo‐sections Observed in Stem

Frederik, PM Peter; Busing, W.M.; Hax, W.M.A.

doi:10.1111/j.1365-2818.1982.tb00352.x

Cited by 9 publications

(5 citation statements)

References 4 publications

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“…Contrast within images of ultrathin frozen-hydrated sections has generally been considered too weak for much biological utility (Zierold et al 1982;Frederik, Busing & Hax, 19826;Hutchinson, Johnson & MacKenzie, 1978;Sasaki et al 1983). But recently McDowall et al (1983) have obtained images of ultrathin frozenhydrated sections of rat liver with clear identification of mitochondria, nuclear and plasma membranes and many other features.…”

Section: Assay Of Chemical Elements By Microprobe Methodsmentioning

confidence: 99%

The localization and assay of chemical elements by microprobe methods

Hall

Gupta

1983

Quart. Rev. Biophys.

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The principle of the microprobe analysis of chemical elements is illustrated in Fig. i. Some kind of radiation is directed on to the specimen, generating signals characteristic of the elements present. Local analysis in situ is achieved in one of two ways. Most often the impinging beam is finely focused so that the signal at any moment comes only from a selected microregion. Alternatively, in some instruments, the impinging beam floods a larger region but the emergent signals characteristic of a particular element may be selected and focused to give an elemental ‘map’ or ‘image’.

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Section: Assay Of Chemical Elements By Microprobe Methodsmentioning

confidence: 99%

The localization and assay of chemical elements by microprobe methods

Hall

Gupta

1983

Quart. Rev. Biophys.

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“…The problems encountered and the results obtained are discussed by Zierold (1982) and Gupta & Hall (1981). Structural observations on frozen hydrated biological materials are incidental and deceptive (Echlin, 1978;Gupta & Hall, 1981 ; Saubermann et al, Heide & Grund, 1974;Hutchinson el al., 1978;Frederik et al, 1982;Zierold, 1982). Contrast is very low and only large structures can be recognized.…”

Section: Introductionmentioning

confidence: 99%

Electron microscopy of frozen hydrated sections of vitreous ice and vitrified biological samples

McDowall

Chang

Freeman

et al. 1983

Journal of Microscopy

268

137

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SUMMARY The preparation and high resolution observation of frozen hydrated thin sections has been studied by transmission electron microscopy (TEM and STEM) on model systems, including pure water, protein solutions, catalase crystals, myelin sheath and various tissues. The state of the ice is determined by electron diffraction. Mass measurement in the electron microscope is used to determine section thickness and control hydration. An adequate depth of vitrified material for sectioning can be obtained from many biological suspensions or untreated tissues. Frozen hydrated sections around 100 nm thick can be produced under optimal conditions from vitreous ice or from vitrified biological samples. Sectioning, transfer and observation in the electron microscope is feasible without alteration of the sample hydration or its initial vitrification. Biological structures can be preserved and observed down to 10 nm. Under favourable working conditions, specimen compression during sectioning and electron beam damage are the factors limiting high resolution observations.

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“…But successful results concerning the recognition of ultrastructural details in frozen-hydrated thin sections are rare (Hall & Gupta, 1982;McDowall et al, 1983;Dubochet et al, 1983). Often, completely frozen-hydrated sections are reported to show no contrast of biological structures unless they are at least partially freeze-dried (Frederik et al, 1982;Hagler & Buja, 1984;Zierold, 1983Zierold, , 1984. In this paper different imaging modes available in STEM are compared with regard to the contrast achieved from ultrathin frozen-hydrated sections of yeast cells.…”

Section: Introductionmentioning

confidence: 99%

Contrast of biological cryosections in scanning transmission electron microscopy

Zierold

1985

Journal of Microscopy

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SUMMARY Ultrathin (70–100 nm thick) frozen‐hydrated cryosections of yeast cells were studied by scanning transmission electron microscopy (STEM). Generally, the contrast is very poor in brightfield and increases slightly by use of the darkfield signal in imaging. The contrast can either be enhanced by selective staining before cryofixation or by mass loss caused either by freeze‐drying or by radiation damage. Partial freeze‐drying by superficial ice sublimation seems to be the most efficient method to increase the contrast in cryosections in the frozen‐hydrated state. The highest contrast in cryosections is achieved after complete freeze‐drying.

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Frozen‐hydrated and Drying Thin Cryo‐sections Observed in Stem

Cited by 9 publications

References 4 publications

The localization and assay of chemical elements by microprobe methods

The localization and assay of chemical elements by microprobe methods

Electron microscopy of frozen hydrated sections of vitreous ice and vitrified biological samples

Contrast of biological cryosections in scanning transmission electron microscopy

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