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Andrew", "lastName": "Hires" }, { "creatorType": "author", "firstName": "Loren L.", "lastName": "Looger" } ], "abstractNote": "Genetically encoded calcium indicators (GECIs), which are based on chimeric fluorescent proteins, can be used to monitor calcium transients in living cells and organisms. Because they are encoded by DNA, GECIs can be delivered to the intact brain noninvasively and targeted to defined populations of neurons and specific subcellular compartments for long-term, repeated measurements in vivo. GECIs have improved iteratively and are becoming useful for imaging neural activity in vivo. Here we summarize extrinsic and intrinsic factors that influence a GECI’s performance and provides guidelines for selecting the appropriate GECI for a given application. 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The model describes intracellular calcium handling under simplified conditions, for which analytical expressions for the amplitude and the time constants of [Ca2+]i changes can be explicitly derived. In particular, it reveals the dependence of the measured [Ca2+]i changes on the calcium indicator concentration. Applied to experimental data from small cells or subcellular compartments, the model equations have been extremely useful for obtaining quantitative information about essential parameters of Ca2+ influx, buffering, and clearance. We illustrate also several changes that occur when the basic assumptions do not hold (e.g., when calcium diffusion, dye saturation, or kinetic effects become significant). 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Understanding the principles underlying fluorescence microscopy is useful when attempting to solve imaging problems. Additionally, fluorescence microscopy is in a state of rapid evolution, with new techniques, probes and equipment appearing almost daily. Familiarity with fluorescence is a prerequisite for taking advantage of many of these developments. This review attempts to provide a framework for understanding excitation of and emission by fluorophores, the way fluorescence microscopes work, and some of the ways fluorescence can be optimized.", "publicationTitle": "Nature Methods", "publisher": "", "place": "", "date": "December 2005", "volume": "2", "issue": "12", "section": "", "partNumber": "", "partTitle": "", "pages": "910-919", "series": "", "seriesTitle": "", "seriesText": "", "journalAbbreviation": "Nat Meth", "DOI": "10.1038/nmeth817", "citationKey": "", "url": "http://www.nature.com/nmeth/journal/v2/n12/abs/nmeth817.html", "accessDate": "2017-02-27T00:43:46Z", "PMID": "", "PMCID": "", "ISSN": "1548-7091", "archive": "", "archiveLocation": "", "shortTitle": "", "language": "en", "libraryCatalog": "www.nature.com", "callNumber": "", "rights": "© 2005 Nature Publishing Group", "extra": "", "tags": [], "collections": [ "T42FBGUC" ], "relations": {}, "dateAdded": "2017-02-27T00:43:46Z", "dateModified": "2017-02-27T00:43:46Z" } }, { "key": "PV672KS6", "version": 47, "library": { "type": "group", "id": 163905, "name": "LeiferLab_OldSchool", "links": { "alternate": { "href": "https://www.zotero.org/groups/leiferlab_oldschool", "type": "text/html" } } }, "links": { "self": { "href": "https://api.zotero.org/groups/163905/items/PV672KS6", "type": "application/json" }, "alternate": { "href": "https://www.zotero.org/groups/leiferlab_oldschool/items/PV672KS6", "type": "text/html" }, "up": { "href": "https://api.zotero.org/groups/163905/items/M7I5CJFU", "type": "application/json" } }, "meta": { "createdByUser": { "id": 8015, "username": "Kramer", "name": "Andrew Leifer", "links": { "alternate": { "href": "https://www.zotero.org/kramer", "type": "text/html" } } } }, "data": { "key": "PV672KS6", "version": 47, "parentItem": "M7I5CJFU", "itemType": "attachment", "linkMode": "linked_url", "title": "Full Text PDF", "accessDate": "2017-02-27T00:43:46Z", "url": "http://www.nature.com/nmeth/journal/v2/n12/pdf/nmeth817.pdf", "note": "", "contentType": "application/pdf", "charset": "", "tags": [], "relations": {}, "dateAdded": "2017-02-27T00:43:46Z", "dateModified": "2017-02-27T00:43:46Z" } }, { "key": "8HHS878P", "version": 47, "library": { "type": "group", "id": 163905, "name": "LeiferLab_OldSchool", "links": { "alternate": { "href": "https://www.zotero.org/groups/leiferlab_oldschool", "type": "text/html" } } }, "links": { "self": { "href": "https://api.zotero.org/groups/163905/items/8HHS878P", "type": "application/json" }, "alternate": { "href": "https://www.zotero.org/groups/leiferlab_oldschool/items/8HHS878P", "type": "text/html" }, "up": { "href": "https://api.zotero.org/groups/163905/items/M7I5CJFU", "type": "application/json" } }, "meta": { "createdByUser": { "id": 8015, "username": "Kramer", "name": "Andrew Leifer", "links": { "alternate": { "href": "https://www.zotero.org/kramer", "type": "text/html" } } } }, "data": { "key": "8HHS878P", "version": 47, "parentItem": "M7I5CJFU", "itemType": "attachment", "linkMode": "linked_url", "title": "Snapshot", "accessDate": "2017-02-27T00:43:46Z", "url": "http://www.nature.com/nmeth/journal/v2/n12/abs/nmeth817.html", "note": "", "contentType": "text/html", "charset": "", "tags": [], "relations": {}, "dateAdded": "2017-02-27T00:43:46Z", "dateModified": "2017-02-27T00:43:46Z" } }, { "key": "J3EFVP9P", "version": 50, "library": { "type": "group", "id": 163905, "name": "LeiferLab_OldSchool", "links": { "alternate": { "href": "https://www.zotero.org/groups/leiferlab_oldschool", "type": "text/html" } } }, "links": { "self": { "href": "https://api.zotero.org/groups/163905/items/J3EFVP9P", "type": "application/json" }, "alternate": { "href": "https://www.zotero.org/groups/leiferlab_oldschool/items/J3EFVP9P", "type": "text/html" } }, "meta": { "createdByUser": { "id": 8015, "username": "Kramer", "name": "Andrew Leifer", "links": { "alternate": { "href": "https://www.zotero.org/kramer", "type": "text/html" } } }, "creatorSummary": "Briggman et al.", "parsedDate": "2005-02-11", "numChildren": 3 }, "data": { "key": "J3EFVP9P", "version": 50, "itemType": "journalArticle", "title": "Optical Imaging of Neuronal Populations During Decision-Making", "creators": [ { "creatorType": "author", "firstName": "K. 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Hyperpolarizing cell 208 during a stimulus biases the leech to swim; depolarizing it biases the leech to crawl or to delay swimming.", "publicationTitle": "Science", "publisher": "", "place": "", "date": "02/11/2005", "volume": "307", "issue": "5711", "section": "", "partNumber": "", "partTitle": "", "pages": "896-901", "series": "", "seriesTitle": "", "seriesText": "", "journalAbbreviation": "Science", "DOI": "10.1126/science.1103736", "citationKey": "", "url": "http://www.sciencemag.org/content/307/5711/896", "accessDate": "2014-10-23T20:10:37Z", "PMID": "15705844", "PMCID": "", "ISSN": "0036-8075, 1095-9203", "archive": "", "archiveLocation": "", "shortTitle": "", "language": "en", "libraryCatalog": "www.sciencemag.org", "callNumber": "", "rights": "", "extra": "", "tags": [], "collections": [], "relations": {}, "dateAdded": "2014-10-23T20:10:37Z", "dateModified": "2014-10-23T20:10:37Z" } }, { "key": "QGEZHKJI", "version": 46, "library": { "type": "group", "id": 163905, "name": 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