Ultrastructure Of Cell Organelles By Scanning Electron Microscopy Of Thick Sections Surface‐Etched By An Oxygen Plasma

Humphreys, W. J.; Henk, William G.

doi:10.1111/j.1365-2818.1979.tb00206.x

Cited by 14 publications

(2 citation statements)

References 11 publications

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“…Plasma etching and ashing are techniques used in a range of industrial processes, but in biology are typically applied to visualize mineral elements in tissues (Hohman & Schraer, 1972;Dinsdale et al, 1979;Chapman, 1985;Arnold et al, 1990;Bogren et al, 1995). A higher SEM resolution of etch-resistant cell components of fixed and embedded tissues is often claimed (Humphreys & Henk, 1979;Yang et al, 1988). Initially, the procedure was used to obtain accurate counts of the number of testes in the cestode Atractolytocestus huronensis that were needed for taxonomic purposes.…”

Section: Discussionmentioning

confidence: 99%

Plasma etching and ashing: a technique for demonstrating internal structures of helminths using scanning electron microscopy

Veltkamp

Chubb²

2006

J. Helminthol.

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Plasma etching and ashing for demonstrating the three-dimensional ultrastructure of the internal organs of helminths is described. Adult worms of the cestode Caryophyllaeides fennica were dehydrated through an ethanol series, critical point dried (Polaron E3000) and sputter coated with 60% gold-palladium (Polaron E5100) and glued to a standard scanning electron microscope (SEM) stub positioned as required for ashing. After initial SEM viewing of worm surfaces for orientation, stubs were placed individually in the reactor chamber of a PT7150 plasma etching and ashing machine. Worms were exposed to a radio frequency (RF) potential in a low pressure (0.2 mbar) oxygen atmosphere at room temperature. The oxidation process was controlled by varying the times of exposure to the RF potential between 2 to 30 min, depending on the depth of surface tissue to be removed to expose target organs or tissues. After each exposure the oxidized layer was blown from the surface with compressed air, the specimen sputter-coated, and viewed by SEM. The procedure was repeated as necessary, to progressively expose successive layers. Fine details of organs, cells within, and cell contents were revealed. Ashing has the advantage of providing three dimensional images of the arrangement of organs that are impossible to visualize by any other procedure, for example facilitating testes counts in cestodes. Both freshly-fixed and long-term stored helminths can be ashed. Ashing times to obtain the desired results were determined by trial so that some duplicate material was needed.

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Section: Discussionmentioning

confidence: 99%

Plasma etching and ashing: a technique for demonstrating internal structures of helminths using scanning electron microscopy

Veltkamp

Chubb²

2006

J. Helminthol.

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“…sections also enhances surface structures in scanning electron microscopy (Humphreys and Henk 1979). Etch or ash pattern spatial resolutions of lOnm o r better have been obtained (Thomas 1974).…”

Section: Introductionmentioning

confidence: 97%

Microincineration and elemental X-ray microanalysis of single Bacillus cereus T spores

Scherrer¹,

Shull²

1987

Can. J. Microbiol.

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Single whole spores of bacillus cereus T were analyzed by scanning electron microscopy and electron microprobe X-ray microanalysis before and after high-temperature (600 degrees C) ashing in air. High-temperature ashing consisted of a centripetal oxidation of the spore surface combined with pyrolysis of the spore's interior. Ashing of single spores produced a compact central ash particle, mimicking the much larger unashed spore body in outline but containing craterlike microregions, and a peripheral thin ash film. Ashing mostly eliminated the spore's organic matrix; however, ash residues still gave residual carbon-characteristic X-ray counts. Ashing of single spores produced a two-, five-, and six-fold increase of potassium, magnesium, and calcium X-ray intensities, respectively. Iron, although low in actual counts, became detectable after ashing. Phosphorus characteristic X-rays were decreased by 41% after ashing, while volatilization lowered sodium and manganese X-ray intensities by over 80%. High-temperature ashing enhanced element-characteristic X-ray intensities of the non-volatilizable mineral(ized) elements of spores by compacting them into ash residues, more so than by simply abolishing their organic matrix. Microincineration appears a generally useful preconcentration technique for elemental detection and localization in X-ray microanalysis.

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Steedman's polyester wax embedment and de‐embedment for combined light and scanning electron microscopy

Norenburg

Barrett

1987

J. Elec. Microsc. Tech.

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Georgia 30912 (J. M.B.) KEY WORDSTechnique, SEM, Histology, Polyester wax ABSTRACT Steedman's polyester wax mixture is a good, general-purpose histological embedding medium that is suitable and convenient to use when it is desirable to combine light microscopy with scanning electron microscopy (SEMI. A range of properties recommend this wax: it has a low melting temperature (37"C), is readily soluble in most dehydrating agents, results in negligible tissue shrinkage, preserves tissue antigenicity, and may even be used as a solvent for fixative agents. We prepare and embed tissues in polyester for light microscopy much as they would be for paraffin wax. For SEM, the block surface is micro-or ultraplaned, utilizing, respectively, a standard rotary microtome with razor blade knives or an ultramicrotome with glass knives. The block is de-waxed in absolute alcohol and then taken to critical point drying. Similarly, sections mounted on coverslips or glass slides may be brought to the SEM after removing the wax. This enables one to bring to the SEM relatively large block faces or sections with good control over orientation. We find the results to be superior to similar procedures employing paraffin. We believe it to be more versatile and equivalent or superior to a variety of other techniques designed to gain access to the interior of tissues with SEM.

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Ultrastructure Of Cell Organelles By Scanning Electron Microscopy Of Thick Sections Surface‐Etched By An Oxygen Plasma

Cited by 14 publications

References 11 publications

Plasma etching and ashing: a technique for demonstrating internal structures of helminths using scanning electron microscopy

Plasma etching and ashing: a technique for demonstrating internal structures of helminths using scanning electron microscopy

Microincineration and elemental X-ray microanalysis of single Bacillus cereus T spores

Steedman's polyester wax embedment and de‐embedment for combined light and scanning electron microscopy

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