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Quantitative energy-dispersive X-ray microanalysis of calcium dynamics in cell suspensions during stimulation on a subsecond time scale : preparative and analytical aspects as exemplified with Paramecium Cells

Quantitative energy-dispersive X-ray microanalysis of calcium dynamics in cell suspensions during stimulation on a subsecond time scale : preparative and analytical aspects as exemplified with Paramecium Cells

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HARDT, Martin, Helmut PLATTNER, 1999. Quantitative energy-dispersive X-ray microanalysis of calcium dynamics in cell suspensions during stimulation on a subsecond time scale : preparative and analytical aspects as exemplified with Paramecium Cells. In: Journal of Structural Biology. 128(2), pp. 187-199. ISSN 1047-8477

@article{Hardt1999Quant-8546, title={Quantitative energy-dispersive X-ray microanalysis of calcium dynamics in cell suspensions during stimulation on a subsecond time scale : preparative and analytical aspects as exemplified with Paramecium Cells}, year={1999}, doi={10.1006/jsbi.1999.4188}, number={2}, volume={128}, issn={1047-8477}, journal={Journal of Structural Biology}, pages={187--199}, author={Hardt, Martin and Plattner, Helmut} }

<rdf:RDF xmlns:rdf="http://www.w3.org/1999/02/22-rdf-syntax-ns#" xmlns:bibo="http://purl.org/ontology/bibo/" xmlns:dc="http://purl.org/dc/elements/1.1/" xmlns:dcterms="http://purl.org/dc/terms/" xmlns:xsd="http://www.w3.org/2001/XMLSchema#" > <rdf:Description rdf:about="https://kops.uni-konstanz.de/rdf/resource/123456789/8546"> <dc:contributor>Plattner, Helmut</dc:contributor> <dcterms:title>Quantitative energy-dispersive X-ray microanalysis of calcium dynamics in cell suspensions during stimulation on a subsecond time scale : preparative and analytical aspects as exemplified with Paramecium Cells</dcterms:title> <dcterms:issued>1999</dcterms:issued> <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2011-03-24T17:44:34Z</dcterms:available> <dc:creator>Plattner, Helmut</dc:creator> <dcterms:bibliographicCitation>First publ. in: Journal of Structural Biology ; 128 (1999), 2. - S. 187-199</dcterms:bibliographicCitation> <dcterms:abstract xml:lang="eng">We analyzed preparative and analytical aspects of the dynamic localization of Ca2+ during cell stimulation, using a combination of quenched flow and energy-dispersive X-ray microanalysis (EDX). Calcium (or Sr, as a substitute) was retained as fluorides during freeze-substitution, followed by epoxide embedding. The quenched-flow used allowed analyses, during stimulation, in the subsecond time range. Sections of 500 nm were analyzed and no artificial Ca or Sr leakage was recognizable. We calculated a primary beam spread from 63 to 72 nm that roughly indicated the resolution of EDX/structure correlation. These values are quite compatible with the size of potential structures of interest, e.g., Ca stores (~100-nm thickness) or cilia (~250-nm diameter). We used widely different standards to calibrate the ratio of CaK net counts in relation to actual [Ca]. Calibration curves showed a linear relationship and a detection limit of [Ca] = 2 mM, while [Ca] in cytosol was 3 mM and in stores was 43 mM, both in nonactivated cells. Eventually Sr2+ can rapidly be substituted for Ca2+ in the medium before and during stimulation, thus allowing one to determine Me2+ fluxes. With our ''model'' cell, Paramecium, we showed that, upon stimulation (causing rapid Ca2+ mobilization from subplasmalemmal stores), Ca was immediately exchanged for Sr in stores.</dcterms:abstract> <dcterms:rights rdf:resource="https://creativecommons.org/licenses/by-nc-nd/2.0/legalcode"/> <dc:creator>Hardt, Martin</dc:creator> <bibo:uri rdf:resource="http://kops.uni-konstanz.de/handle/123456789/8546"/> <dc:format>application/pdf</dc:format> <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2011-03-24T17:44:34Z</dc:date> <dc:language>eng</dc:language> <dc:contributor>Hardt, Martin</dc:contributor> <dc:rights>deposit-license</dc:rights> </rdf:Description> </rdf:RDF>

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