Quantifying Exocytosis by Combination of Membrane Capacitance Measurements and Total Internal Reflection Fluorescence Microscopy in Chromaffin Cells

Hdl Handle:
http://hdl.handle.net/10675.2/535
Title:
Quantifying Exocytosis by Combination of Membrane Capacitance Measurements and Total Internal Reflection Fluorescence Microscopy in Chromaffin Cells
Authors:
Becherer, Ute ( 0000-0001-6005-7517 ) ; Pasche, Mathias; Nofal, Shahira; Hof, Detlef; Matti, Ulf; Rettig, Jens
Abstract:
Total internal reflection fluorescence microscopy (TIRF-Microscopy) allows the observation of individual secretory vesicles in real-time during exocytosis. In contrast to electrophysiological methods, such as membrane capacitance recording or carbon fiber amperometry, TIRF-Microscopy also enables the observation of vesicles as they reside close to the plasma membrane prior to fusion. However, TIRF-Microscopy is limited to the visualization of vesicles that are located near the membrane attached to the glass coverslip on which the cell grows. This has raised concerns as to whether exocytosis measured with TIRF-Microscopy is comparable to global secretion of the cell measured with membrane capacitance recording. Here we address this concern by combining TIRF-Microscopy and membrane capacitance recording to quantify exocytosis from adrenal chromaffin cells. We found that secretion measured with TIRF-Microscopy is representative of the overall secretion of the cells, thereby validating for the first time the TIRF method as a measure of secretion. Furthermore, the combination of these two techniques provides a new tool for investigating the molecular mechanism of synaptic transmission with combined electrophysiological and imaging techniques.
Editors:
Mei, Lin
Citation:
PLoS ONE. 2007 Jun 6; 2(6):e505
Issue Date:
6-Jun-2007
URI:
http://hdl.handle.net/10675.2/535
DOI:
10.1371/journal.pone.0000505
PubMed ID:
17551585
PubMed Central ID:
PMC1876815
Type:
Article
ISSN:
1932-6203
Appears in Collections:
Department of Neurology: Faculty Research and Presentations

Full metadata record

DC FieldValue Language
dc.contributor.authorBecherer, Uteen_US
dc.contributor.authorPasche, Mathiasen_US
dc.contributor.authorNofal, Shahiraen_US
dc.contributor.authorHof, Detlefen_US
dc.contributor.authorMatti, Ulfen_US
dc.contributor.authorRettig, Jensen_US
dc.contributor.editorMei, Lin-
dc.date.accessioned2012-10-26T16:26:33Z-
dc.date.available2012-10-26T16:26:33Z-
dc.date.issued2007-06-6en_US
dc.identifier.citationPLoS ONE. 2007 Jun 6; 2(6):e505en_US
dc.identifier.issn1932-6203en_US
dc.identifier.pmid17551585en_US
dc.identifier.doi10.1371/journal.pone.0000505en_US
dc.identifier.urihttp://hdl.handle.net/10675.2/535-
dc.description.abstractTotal internal reflection fluorescence microscopy (TIRF-Microscopy) allows the observation of individual secretory vesicles in real-time during exocytosis. In contrast to electrophysiological methods, such as membrane capacitance recording or carbon fiber amperometry, TIRF-Microscopy also enables the observation of vesicles as they reside close to the plasma membrane prior to fusion. However, TIRF-Microscopy is limited to the visualization of vesicles that are located near the membrane attached to the glass coverslip on which the cell grows. This has raised concerns as to whether exocytosis measured with TIRF-Microscopy is comparable to global secretion of the cell measured with membrane capacitance recording. Here we address this concern by combining TIRF-Microscopy and membrane capacitance recording to quantify exocytosis from adrenal chromaffin cells. We found that secretion measured with TIRF-Microscopy is representative of the overall secretion of the cells, thereby validating for the first time the TIRF method as a measure of secretion. Furthermore, the combination of these two techniques provides a new tool for investigating the molecular mechanism of synaptic transmission with combined electrophysiological and imaging techniques.en_US
dc.rightsBecherer et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.en_US
dc.subjectResearch Articleen_US
dc.subjectCell Biology/Membranes and Sortingen_US
dc.subjectNeuroscience/Neuronal and Glial Cell Biologyen_US
dc.subjectNeuroscience/Neuronal Signaling Mechanismsen_US
dc.titleQuantifying Exocytosis by Combination of Membrane Capacitance Measurements and Total Internal Reflection Fluorescence Microscopy in Chromaffin Cellsen_US
dc.typeArticleen_US
dc.identifier.pmcidPMC1876815en_US
dc.contributor.corporatenameDepartment of Neurology-
dc.contributor.corporatenameCollege of Graduate Studies-

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