Silver‐enhanced colloidal‐gold labelling of rabbit kidney collecting‐duct cell surfaces imaged by scanning electron microscopy

Herter, Peter; Laube, Gregor; Gronczewski, J.; Minuth, Will W.

doi:10.1111/j.1365-2818.1993.tb03364.x

Cited by 14 publications

(13 citation statements)

References 37 publications

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“…From previous experiments we knew that aldosterone can modulate differentiation [22] and increase the number of cells which bind PNA on their luminal plasma membrane, as do β-type IC cells in the adult kidney [16,29,30]. It is not known if aldosterone acts on the embryonic epithelia on the basolateral side as it does in the adult organ, or if there is a luminal effect during organ maturation.…”

Section: Resultsmentioning

confidence: 93%

Culture of embryonic renal collecting duct epithelia in a gradient container

Minuth

Aigner

Kloth

et al. 1997

Pediatric Nephrology

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During organogenesis the ampullar epithelium of the renal collecting duct acts as an inducer which generates all of the nephron anlagen. As development proceeds, one part of the collecting duct cells in the ampullar tip retain their inducer capability, while others develop into the functional epithelium consisting of principal and intercalated (IC) cells. The events leading from the embryonic inducer to the mature tissue are unknown. We investigated the maturation of embryonic collecting duct epithelium derived from neonatal rabbit kidney under in vitro conditions. To prevent dedifferentiation the epithelia were cultured on kidney-specific support material within a tissue carrier. Apical and basal compartments of the epithelia were simulated in a gradient culture container. The two sides of the epithelium were each constantly perfused with a different medium. During the 14-day incubation the tissue was not subcultured. The development of collecting duct cell features was investigated with morphological and immunohistochemical methods. Both light and electron microscopy revealed morphologically intact epithelia following gradient culture. The polarized cells rested on a uniformly developed basement membrane. The continuous application of aldosterone during the culture modulated the development of collecting duct cell characteristics. Both basal and luminal administration of aldosterone initiated differentiation in the embryonic epithelia. Using the sodium (Na) channel blocker amiloride, it was demonstrated that Na channels are involved in the differentiation of the IC cell phenotype.

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Section: Resultsmentioning

confidence: 93%

Culture of embryonic renal collecting duct epithelia in a gradient container

Minuth

Aigner

Kloth

et al. 1997

Pediatric Nephrology

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“…A slight degree of background silver deposits was a feature of some experiments; however, this is a wellrecognised complication of this method (Herter et al 1993). …”

Section: Controlsmentioning

confidence: 96%

“…Following further washes (PBS/0.5% BSA), tissues were incubated in goat anti-mouse IgG-1-nm gold conjugate (British Biocell International) and then washed in PBS followed by distilled water. The tissues were then fixed in 2% glutaraldehyde prior to silver enhancement (British Biocell International; Herter et al 1993). A final wash in distilled water preceded dehydration and critical-point drying.…”

Section: Immunohistochemistrymentioning

confidence: 99%

“…The tissues can be imaged in a gaseous environment in the specimen chamber in the backscattered electron mode to provide strong material contrast (Danilatos 1993;Griffin 2000). As there is no need for the conductive coating of biological material, immunophenotypic characterisation of cells can be achieved by staining with colloidal-gold conjugated immunoreagents followed by silver enhancement (Herter et al 1993). Immunopositive cells are then readily detected by ESEM (because of the silver staining) in the context of the surrounding surface tissue microarchitecture.…”

Section: Introductionmentioning

confidence: 99%

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Macrophages and dendritic cells in the rat meninges and choroid plexus: three-dimensional localisation by environmental scanning electron microscopy and confocal microscopy

McMenamin

Wealthall

Deverall

et al. 2003

Cell and Tissue Research

135

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The present investigation provides novel information on the topographical distribution of macrophages and dendritic cells (DCs) in normal meninges and choroid plexus of the rat central nervous system (CNS). Whole-mounts of meninges and choroid plexus of Lewis rats were incubated with various anti-leucocyte monoclonal antibodies and either visualised with gold-conjugated secondary antibody followed by silver enhancement and subsequent examination by environmental scanning electron microscopy or by the use of fluorochromes and confocal microscopy. Large numbers of MHC class II(+) putative DCs were identified on the internal or subarachnoid aspect of dural whole-mounts, on the surface of the cortex (pia/arachnoid) and on the surface of the choroid plexus. Occupation of these sites would allow DCs access to cerebrospinal fluid (CSF) and therefore allow antigens into the subarachnoid space and ventricles. By contrast, macrophages were less evident at sites exposed to CSF and were more frequently located within the connective tissue of the dura/arachnoid and choroid plexus stroma and also in a sub-pial location. The present data suggest that DC may be strategically located within the CNS to sample CSF-borne antigens. Furthermore, the data suggest that CNS tissue samples collected without careful removal of the meninges may inadvertently be contaminated by DCs and meningeal macrophages.

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“…In high vacuum microscopy with chromium-coated cells, SE mode failed to identify gold labels while the BSE signal, because of the great difference of atomic number between chromium and gold, produced a contrast sufficient to correlate the distribution of gold particles to the morphological details of the cell surface (Herter et al, 1993). Coating with carbon offers less contrast than chromium, so it maximizes gold evidence but hides cell surface details.…”

Section: Esem Biological Applicationsmentioning

confidence: 99%

A critical overview of ESEM applications in the biological field

Muscariello

Rosso

Marino

et al. 2005

Journal Cellular Physiology

187

116

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Scanning Electron Microscope (SEM) is a powerful research tool, but since it requires high vacuum conditions, the wet materials and biological samples must undergo a complex preparation that limits the application of SEM on this kind of specimen and often causes the introduction of artifacts. The introduction of Environmental Scanning Electron Microscope (ESEM), working in gaseous atmosphere, represented a new perspective in biological research. Despite the fact that many biological applications have demonstrated the convenience of ESEM, the full potentialities of this technology are still under investigation. In this review, the exploration of the recent literature data confronted with the first results obtained in our experimental work suggest that ESEM represents an important extension of conventional scanning microscopy.

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Silver‐enhanced colloidal‐gold labelling of rabbit kidney collecting‐duct cell surfaces imaged by scanning electron microscopy

Cited by 14 publications

References 37 publications

Culture of embryonic renal collecting duct epithelia in a gradient container

Culture of embryonic renal collecting duct epithelia in a gradient container

Macrophages and dendritic cells in the rat meninges and choroid plexus: three-dimensional localisation by environmental scanning electron microscopy and confocal microscopy

A critical overview of ESEM applications in the biological field

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