Detection of Secretion from Single Pancreatic β-Cells Using Extracellular Fluorogenic Reactions and Confocal Fluorescence Microscopy
Confocal microscopy with Zinquin, a fluorogenic Zn2+-specific indicator, was used for spatially and temporally resolved measurement of Zn2+ efflux from single pancreatic β-cells. When cells were incubated in buffer containing Zinquin, application of insulin secretagogues evoked an increase in fluore...
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| Veröffentlicht in: | Analytical chemistry (Washington) 2000-02, Vol.72 (4), p.711-717 |
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| creator | Qian, Wei-Jun Aspinwall, Craig A Battiste, Merle A Kennedy, Robert T |
| description | Confocal microscopy with Zinquin, a fluorogenic Zn2+-specific indicator, was used for spatially and temporally resolved measurement of Zn2+ efflux from single pancreatic β-cells. When cells were incubated in buffer containing Zinquin, application of insulin secretagogues evoked an increase in fluorescence around the surface of the cell, indicative of detection of Zn2+ efflux from the cell. The fluorescence increases corresponded spatially and temporally with measurements of exocytosis obtained simultaneously by amperometry. When images were taken at 266-ms intervals, the detection limit for Zn2+ was ∼0.5 μM. With this image frequency, it was possible to observe bursts of fluorescence which were interpreted as fluctuations of Zn2+ level due to exocytosis. The average intensity of these fluorescence bursts corresponded to a Zn2+ concentration of ∼7 μM. Since insulin is co-stored with Zn2+ in secretory vesicles, it was concluded that the Zn2+ efflux corresponded to exocytosis of insulin/Zn2+-containing granules from the β-cell. Exocytosis sites identified by this technique were frequently localized to one portion of the cell, indicative of active areas of release. |
| doi_str_mv | 10.1021/ac991085t |
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When cells were incubated in buffer containing Zinquin, application of insulin secretagogues evoked an increase in fluorescence around the surface of the cell, indicative of detection of Zn2+ efflux from the cell. The fluorescence increases corresponded spatially and temporally with measurements of exocytosis obtained simultaneously by amperometry. When images were taken at 266-ms intervals, the detection limit for Zn2+ was ∼0.5 μM. With this image frequency, it was possible to observe bursts of fluorescence which were interpreted as fluctuations of Zn2+ level due to exocytosis. The average intensity of these fluorescence bursts corresponded to a Zn2+ concentration of ∼7 μM. Since insulin is co-stored with Zn2+ in secretory vesicles, it was concluded that the Zn2+ efflux corresponded to exocytosis of insulin/Zn2+-containing granules from the β-cell. Exocytosis sites identified by this technique were frequently localized to one portion of the cell, indicative of active areas of release.</description><identifier>ISSN: 0003-2700</identifier><identifier>EISSN: 1520-6882</identifier><identifier>DOI: 10.1021/ac991085t</identifier><identifier>PMID: 10701254</identifier><identifier>CODEN: ANCHAM</identifier><language>eng</language><publisher>Washington, DC: American Chemical Society</publisher><subject>Animals ; Biological and medical sciences ; Cell physiology ; Cells, Cultured ; Cellular biology ; Chemistry ; Exocytosis ; Extracellular Space - chemistry ; Fluorescent Dyes ; Fundamental and applied biological sciences. Psychology ; Intracellular Fluid - chemistry ; Islets of Langerhans - chemistry ; Islets of Langerhans - metabolism ; Mice ; Microscopy, Confocal ; Microscopy, Fluorescence ; Molecular and cellular biology ; Pancreas ; Quinolones ; Secretion. Exocytosis ; Tosyl Compounds ; Zinc - analysis ; Zinc - metabolism</subject><ispartof>Analytical chemistry (Washington), 2000-02, Vol.72 (4), p.711-717</ispartof><rights>Copyright © 2000 American Chemical Society</rights><rights>2001 INIST-CNRS</rights><rights>Copyright American Chemical Society Feb 15, 2000</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a404t-e1ed78c7fe41bd7c4faf65e9021f8355ada85283f367b361fa99cc0900c288303</citedby><cites>FETCH-LOGICAL-a404t-e1ed78c7fe41bd7c4faf65e9021f8355ada85283f367b361fa99cc0900c288303</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/ac991085t$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/ac991085t$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>314,780,784,2763,27075,27923,27924,56737,56787</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=811651$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10701254$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Qian, Wei-Jun</creatorcontrib><creatorcontrib>Aspinwall, Craig A</creatorcontrib><creatorcontrib>Battiste, Merle A</creatorcontrib><creatorcontrib>Kennedy, Robert T</creatorcontrib><title>Detection of Secretion from Single Pancreatic β-Cells Using Extracellular Fluorogenic Reactions and Confocal Fluorescence Microscopy</title><title>Analytical chemistry (Washington)</title><addtitle>Anal. Chem</addtitle><description>Confocal microscopy with Zinquin, a fluorogenic Zn2+-specific indicator, was used for spatially and temporally resolved measurement of Zn2+ efflux from single pancreatic β-cells. When cells were incubated in buffer containing Zinquin, application of insulin secretagogues evoked an increase in fluorescence around the surface of the cell, indicative of detection of Zn2+ efflux from the cell. The fluorescence increases corresponded spatially and temporally with measurements of exocytosis obtained simultaneously by amperometry. When images were taken at 266-ms intervals, the detection limit for Zn2+ was ∼0.5 μM. With this image frequency, it was possible to observe bursts of fluorescence which were interpreted as fluctuations of Zn2+ level due to exocytosis. The average intensity of these fluorescence bursts corresponded to a Zn2+ concentration of ∼7 μM. Since insulin is co-stored with Zn2+ in secretory vesicles, it was concluded that the Zn2+ efflux corresponded to exocytosis of insulin/Zn2+-containing granules from the β-cell. Exocytosis sites identified by this technique were frequently localized to one portion of the cell, indicative of active areas of release.</description><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Cell physiology</subject><subject>Cells, Cultured</subject><subject>Cellular biology</subject><subject>Chemistry</subject><subject>Exocytosis</subject><subject>Extracellular Space - chemistry</subject><subject>Fluorescent Dyes</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Intracellular Fluid - chemistry</subject><subject>Islets of Langerhans - chemistry</subject><subject>Islets of Langerhans - metabolism</subject><subject>Mice</subject><subject>Microscopy, Confocal</subject><subject>Microscopy, Fluorescence</subject><subject>Molecular and cellular biology</subject><subject>Pancreas</subject><subject>Quinolones</subject><subject>Secretion. Exocytosis</subject><subject>Tosyl Compounds</subject><subject>Zinc - analysis</subject><subject>Zinc - metabolism</subject><issn>0003-2700</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpl0d1u0zAUB3ALgVgZXPACyAKBxEXg2I5j5xKVbSANqNrt2nKd4ykjjTs7kbYH4IV4EJ4Jd6kKgivL9k9_nQ9CnjN4x4Cz99bVNQMthwdkxiSHotKaPyQzABAFVwBH5ElK1wCMAasekyMGChiX5Yz8-IgDuqENPQ2ertBFvL_4GDZ01fZXHdKF7fOzHVpHf_0s5th1iV6m_EdPbodoXX4YOxvpaTeGGK6wz3CJ9j41Uds3dB56H5ztJoLJYe-QfmldDMmF7d1T8sjbLuGz_XlMLk9PLuafivNvZ5_nH84LW0I5FMiwUdopjyVbN8qV3vpKYp1n4LWQ0jZWS66FF5Vai4p5W9fOQQ3guNYCxDF5M-VuY7gZMQ1m06Zd_bbHMCajoC5FDsrw5T_wOoyxz7UZzlTOkopn9HZCuzZSRG-2sd3YeGcYmN1izGEx2b7YB47rDTZ_yWkTGbzaA5vypHzMQ2_TwWnGKsmyKibVpgFvD782fjeVEkqai8XKfF2IheJnS7PM_vXkrUt_Wvi_vN_SarIA</recordid><startdate>20000215</startdate><enddate>20000215</enddate><creator>Qian, Wei-Jun</creator><creator>Aspinwall, Craig A</creator><creator>Battiste, Merle A</creator><creator>Kennedy, Robert T</creator><general>American Chemical Society</general><scope>BSCLL</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QF</scope><scope>7QO</scope><scope>7QQ</scope><scope>7SC</scope><scope>7SE</scope><scope>7SP</scope><scope>7SR</scope><scope>7TA</scope><scope>7TB</scope><scope>7TM</scope><scope>7U5</scope><scope>7U7</scope><scope>7U9</scope><scope>8BQ</scope><scope>8FD</scope><scope>C1K</scope><scope>F28</scope><scope>FR3</scope><scope>H8D</scope><scope>H8G</scope><scope>H94</scope><scope>JG9</scope><scope>JQ2</scope><scope>KR7</scope><scope>L7M</scope><scope>L~C</scope><scope>L~D</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20000215</creationdate><title>Detection of Secretion from Single Pancreatic β-Cells Using Extracellular Fluorogenic Reactions and Confocal Fluorescence Microscopy</title><author>Qian, Wei-Jun ; Aspinwall, Craig A ; Battiste, Merle A ; Kennedy, Robert T</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a404t-e1ed78c7fe41bd7c4faf65e9021f8355ada85283f367b361fa99cc0900c288303</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Cell physiology</topic><topic>Cells, Cultured</topic><topic>Cellular biology</topic><topic>Chemistry</topic><topic>Exocytosis</topic><topic>Extracellular Space - chemistry</topic><topic>Fluorescent Dyes</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Intracellular Fluid - chemistry</topic><topic>Islets of Langerhans - chemistry</topic><topic>Islets of Langerhans - metabolism</topic><topic>Mice</topic><topic>Microscopy, Confocal</topic><topic>Microscopy, Fluorescence</topic><topic>Molecular and cellular biology</topic><topic>Pancreas</topic><topic>Quinolones</topic><topic>Secretion. Exocytosis</topic><topic>Tosyl Compounds</topic><topic>Zinc - analysis</topic><topic>Zinc - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Qian, Wei-Jun</creatorcontrib><creatorcontrib>Aspinwall, Craig A</creatorcontrib><creatorcontrib>Battiste, Merle A</creatorcontrib><creatorcontrib>Kennedy, Robert T</creatorcontrib><collection>Istex</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Aluminium Industry Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>Ceramic Abstracts</collection><collection>Computer and Information Systems Abstracts</collection><collection>Corrosion Abstracts</collection><collection>Electronics & Communications Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Materials Business File</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ANTE: Abstracts in New Technology & Engineering</collection><collection>Engineering Research Database</collection><collection>Aerospace Database</collection><collection>Copper Technical Reference Library</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Materials Research Database</collection><collection>ProQuest Computer Science Collection</collection><collection>Civil Engineering Abstracts</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Computer and Information Systems Abstracts Academic</collection><collection>Computer and Information Systems Abstracts Professional</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Analytical chemistry (Washington)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Qian, Wei-Jun</au><au>Aspinwall, Craig A</au><au>Battiste, Merle A</au><au>Kennedy, Robert T</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Detection of Secretion from Single Pancreatic β-Cells Using Extracellular Fluorogenic Reactions and Confocal Fluorescence Microscopy</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. Chem</addtitle><date>2000-02-15</date><risdate>2000</risdate><volume>72</volume><issue>4</issue><spage>711</spage><epage>717</epage><pages>711-717</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><coden>ANCHAM</coden><abstract>Confocal microscopy with Zinquin, a fluorogenic Zn2+-specific indicator, was used for spatially and temporally resolved measurement of Zn2+ efflux from single pancreatic β-cells. When cells were incubated in buffer containing Zinquin, application of insulin secretagogues evoked an increase in fluorescence around the surface of the cell, indicative of detection of Zn2+ efflux from the cell. The fluorescence increases corresponded spatially and temporally with measurements of exocytosis obtained simultaneously by amperometry. When images were taken at 266-ms intervals, the detection limit for Zn2+ was ∼0.5 μM. With this image frequency, it was possible to observe bursts of fluorescence which were interpreted as fluctuations of Zn2+ level due to exocytosis. The average intensity of these fluorescence bursts corresponded to a Zn2+ concentration of ∼7 μM. Since insulin is co-stored with Zn2+ in secretory vesicles, it was concluded that the Zn2+ efflux corresponded to exocytosis of insulin/Zn2+-containing granules from the β-cell. Exocytosis sites identified by this technique were frequently localized to one portion of the cell, indicative of active areas of release.</abstract><cop>Washington, DC</cop><pub>American Chemical Society</pub><pmid>10701254</pmid><doi>10.1021/ac991085t</doi><tpages>7</tpages></addata></record> |
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| subjects | Animals Biological and medical sciences Cell physiology Cells, Cultured Cellular biology Chemistry Exocytosis Extracellular Space - chemistry Fluorescent Dyes Fundamental and applied biological sciences. Psychology Intracellular Fluid - chemistry Islets of Langerhans - chemistry Islets of Langerhans - metabolism Mice Microscopy, Confocal Microscopy, Fluorescence Molecular and cellular biology Pancreas Quinolones Secretion. Exocytosis Tosyl Compounds Zinc - analysis Zinc - metabolism |
| title | Detection of Secretion from Single Pancreatic β-Cells Using Extracellular Fluorogenic Reactions and Confocal Fluorescence Microscopy |
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