EELS data acquisition, processing and display for the Zeiss CEM 902 based on lotus 1–2‐3®: application examples from a biological system and inorganic transition metal compounds

Drechsler, Markus; Cantow, H.‐J.

doi:10.1111/j.1365-2818.1991.tb03116.x

Cited by 6 publications

(4 citation statements)

References 27 publications

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“…Spectra were recorded in the spectrum mode of the CEM 902 using an objective aperture of 90 pm and a spectrometer aperture of 100 pm. Spectra were recorded at a magnification of 50,000 unless stated otherwise, so that the recorded area (0.044 pm2 according to Drechsler & Cantow, 1991) was occupied completely by the region of interest. Zero-loss image of conventionally prepared porcine lung parenchyma which was fixed by immersion in osmium tetroxide-glutaraldehyde, post-fixed with osmium tetroxide, stained en bloc with aqueous, halfsaturated uranyl acetate, dehydrated through a graded series of alcohols, transferred to Araldite via propylene oxide and embedded in Araldite.…”

Section: A T E R I a L S A N D M E T H O D Smentioning

confidence: 99%

Electron spectroscopic study (ESI, EELS) of Nanoplast‐embedded mammalian lung

Fehrenbach

Richter

Schnabel

1992

Journal of Microscopy

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SUMMARY The potential of Nanoplast melamine resin embedding for the study of mammalian lung parenchyma was examined by means of electron spectroscopic imaging (ESI) and electron energy‐loss spectroscopy (EELS). Samples were either fixed with glutaralde‐hyde‐paraformaldehyde or glutaraldehyde‐tannic acid, or were directly transferred to the embedding medium without prior fixation. Organic dehydrants, as well as fixatives containing heavy metals and stains, were omitted. A very high level of ultrastructural detail of chromatin, ribosomes, mitochondria and plasma membranes was achieved by ESI from the Nanoplast‐embedded samples. The most prominent gain in ultrastructural detail was achieved when moving from an energy loss just below the L2,3 edge of phosphorus at 132 eV to an energy loss just beyond this edge. This reflects the prominent P L2,3 edge observed by EELS of Nanoplast‐embedded samples in comparison with conventionally processed samples. Thus, taking into account possible sectioning artefacts, excellent heterochromatin images which rely on the phosphorus distribution can be obtained from Nanoplast‐embedded samples by computer‐assisted analysis of electron spectroscopic images. In this respect glutaraldehyde‐paraformaldehyde fixation is preferable to glutaraldehyde‐tannic acid fixation because the presence of silicon, revealed by EELS, in tannic‐acid‐fixed samples may introduce artefacts in phosphorus distribution images obtained by the three‐window method because of the close proximity of the L2,3 edges of silicon and phosphorus.

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Section: A T E R I a L S A N D M E T H O D Smentioning

confidence: 99%

Electron spectroscopic study (ESI, EELS) of Nanoplast‐embedded mammalian lung

Fehrenbach

Richter

Schnabel

1992

Journal of Microscopy

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“…using the minimal dose focusing device. The cumulative specimen dose was calculated according to Drechsler & Cantow (1991) as ≈ 0.7 ke nm −2 for a primary microscopical magnification of 20 000 and 1.4 ke nm −2 for a magnification of 30 000. Representative micrographs of the different liposome preparations are shown below.…”

Section: Methodsmentioning

confidence: 99%

Energy‐filtered cryotransmission electron microscopy of liposomes prepared from human stratum corneum lipids

Schmidtgen

Drechsler

Lasch

et al. 1998

Journal of Microscopy

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We used cryo-TEM to examine the morphology of vesicles formed from lipids of the human stratum corneum (hSC). Human stratum corneum lipid liposomes (hSCLLs) were prepared in buffer at various pH values, using different preparation methods (film method, extrusion, ultrasonication, detergent dialysis). The morphology of hSCLLs at pH 7.4 differed markedly from that of liposomes formed by phospholipids, showing folds, stacks and membrane thickening. At pH 5.0, corresponding to natural conditions at the skin surface, membrane structures are essentially the same as those prepared at pH 7.4. Sharp edges in hSCLLs, branching membranes and stable membrane stacks were explained by the presence of ceramides, the major components and structural elements of human stratum corneum lipids (hSCLs). Thickened areas in the membranes may be caused by the local accumulation of triacylglycerols and cholesterol esters in the hydrophobic interior of the bilayer.

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“…As other cations might also have been precipitated by the applied potassium antimonate in the present investigation, the element composition of the formed precipitates was determined by electron energy-loss spectroscopy (EELS). Electron energy-loss spectra were taken by using an integrated serial detector connected via an interface to an XT personal computer as described elsewhere (Drechsler and Cantow 1991).…”

Section: Electron Energy-loss Spectroscopymentioning

confidence: 99%

Possible Ca 2+ -dependent mechanism of apical outer hair cell modulation within the cochlea of the guinea pig

Heinrich

Maurer

Mann

1998

Cell and Tissue Research

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Calcium ions were precipitated with potassium antimonate after injection of the inorganic calcium channel blocker MnCl2 or the inorganic potassium channel blockers BaCl2 or CsCl into the perilymph of the scala vestibuli of the guinea pig. The spatial distribution of the formed histochemical reaction products within the organ of Corti was studied by energy-filtering transmission-electron microscopy. Compared with untreated control ears, the number of the formed precipitates drastically increased at the extracellular side of the lamina reticularis after application of the various inorganic channel blockers. The apical side of the outer hair cells and the intervening Deiter cells were covered by a thick layer of calcium precipitates, whereas the number of histochemical reaction products was clearly reduced in the nearby acellular tectorial membrane. The high calcium content within the formed reaction products at the lamina reticularis could be demonstrated by elemental mapping and by electron energy-loss spectroscopy. To ascertain the alterations in the amounts of the calcium precipitates within the tectorial membrane after application of the various inorganic channel blockers, the precipitate densities were determined semiquantitatively by an image processing system and the values obtained compared with those of untreated control specimens. The observed histochemical results are in good agreement with published electrophysiological findings concerning the spatial distribution of ion channels located at the apical outer hair cell membrane. The detected alterations in the spatial distribution of calcium precipitates might correspond to calcium-dependent processes involved in outer hair cell modulation.

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EELS data acquisition, processing and display for the Zeiss CEM 902 based on lotus 1–2‐3®: application examples from a biological system and inorganic transition metal compounds

Cited by 6 publications

References 27 publications

Electron spectroscopic study (ESI, EELS) of Nanoplast‐embedded mammalian lung

Electron spectroscopic study (ESI, EELS) of Nanoplast‐embedded mammalian lung

Energy‐filtered cryotransmission electron microscopy of liposomes prepared from human stratum corneum lipids

Possible Ca 2+ -dependent mechanism of apical outer hair cell modulation within the cochlea of the guinea pig

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