Optical stimulation of quantal exocytosis on transparent microchips

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Optical stimulation of quantal exocytosis on transparent microchips

Please use this identifier to cite or link to this item: http://hdl.handle.net/10355/4890

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dc.contributor.advisor Gillis, Kevin D. en
dc.contributor.author Chen, Xiaohui, 1973- en_US
dc.date.accessioned 2010-01-12T18:44:29Z
dc.date.available 2010-01-12T18:44:29Z
dc.date.issued 2007 en_US
dc.date.submitted 2007 Fall en
dc.identifier.other ChenX-092607-D8469 en_US
dc.identifier.uri http://hdl.handle.net/10355/4890
dc.description The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. en_US
dc.description Title from title screen of research.pdf file (viewed on January 30, 2008) en_US
dc.description Includes bibliographical references. en_US
dc.description Vita. en_US
dc.description Thesis (Ph. D.) University of Missouri-Columbia 2007. en_US
dc.description Dissertations, Academic -- University of Missouri--Columbia -- Biological engineering. en_US
dc.description.abstract Photorelease of caged Ca²⁺ is a uniquely powerful tool to study the dynamics of Ca²⁺-triggered exocytosis from individual cells. Using photolithography and other microfabrication techniques, we have developed transparent microchip devices to enable photorelease of caged Ca²⁺ together with electrochemical detection of quantal catecholamine secretion from individual cells or cell arrays as a step towards developing high-throughput experimental devices. A 110 nm - thick transparent Indium-Tin-Oxide (ITO) film was sputter-deposited onto glass coverslips, which were then patterned into 24 cell-sized working electrodes (2̃0 [mu]m by 20 [mu]m). We loaded bovine chromaffin cells with acetoxymethyl (AM) ester derivatives of the Ca²⁺ cage NP-EGTA and Ca²⁺ indicator dye Fura-4F, then transferred these cells onto the working ITO electrodes for amperometric recordings. Upon flash photorelease of caged Ca²+С uniform rise of [Ca²⁺]i within the target cell leads to quantal release of oxidizable catecholamines measured amperometrically by the underlying ITO electrode. We observed a burst of amperometric spikes upon rapid elevation of [Ca²⁺]i and a "priming" effect of sub-stimulatory [Ca²⁺]i on the response of cells to subsequent [Ca²⁺]i elevation, similar to previous reports using different techniques. We conclude that UV photolysis of caged Ca²⁺ is a suitable stimulation technique for higher-throughput studies of Ca²⁺-dependent exocytosis on transparent electrochemical microelectrode arrays. en_US
dc.language.iso en_US en_US
dc.publisher University of Missouri--Columbia en_US
dc.subject.lcsh Exocytosis en_US
dc.subject.lcsh Chromaffin cells en_US
dc.subject.lcsh Calcium ions en_US
dc.subject.lcsh Photolithography en_US
dc.subject.lcsh Microfabrication en_US
dc.subject.lcsh Group 13 elements en_US
dc.subject.lcsh Group 14 elements en_US
dc.title Optical stimulation of quantal exocytosis on transparent microchips en_US
dc.type Thesis en_US
thesis.degree.discipline Biological engineering en_US
thesis.degree.grantor University of Missouri--Columbia en_US
thesis.degree.name Ph. D. en_US
thesis.degree.level Doctoral en_US
dc.identifier.merlin .b61978334 en_US
dc.identifier.oclc 191702789 en_US
dc.relation.ispartofcommunity University of Missouri-Columbia. Graduate School. Theses and Dissertations. Dissertations. 2007 Dissertations
dc.relation.ispartofcollection 2007 Freely available dissertations (MU)


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