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dc.contributor.authorLeeson, Hannah C
dc.contributor.authorChan-Ling, Tailoi
dc.contributor.authorLovelace, Michael D
dc.contributor.authorBrownlie, Jeremy D
dc.contributor.authorWeible, Michael W
dc.contributor.authorGu, Ben J
dc.date.accessioned2020-02-21T04:51:29Z
dc.date.available2020-02-21T04:51:29Z
dc.date.issued2019
dc.identifier.issn1940-087X
dc.identifier.doi10.3791/59313
dc.identifier.urihttp://hdl.handle.net/10072/391779
dc.description.abstractLive-cell flow cytometry is increasingly used among cell biologists to quantify biological processes in a living cell culture. This protocol describes a method whereby live-cell flow cytometry is extended upon to analyze the multiple functions of P2X7 receptor activation in real-time. Using a time module installed on a flow cytometer, live-cell functionality can be assessed and plotted over a given time period to explore the kinetics of calcium influx, transmembrane pore formation, and phagocytosis. This simple method is advantageous as all three canonical functions of the P2X7 receptor can be assessed using one machine, and the gathered data plotted over time provides information on the entire live-cell population rather than single-cell recordings often obtained using technically challenging patch-clamp methods. Calcium influx experiments use a calcium indicator dye, while P2X7 pore formation assays rely on ethidium bromide being allowed to pass through the transmembrane pore formed upon high agonist concentrations. Yellow-green (YG) latex beads are utilized to measure phagocytosis. Specific agonists and antagonists are applied to investigate the effects of P2X7 receptor activity. Individually, these methods can be modified to provide quantitative data on any number of calcium channels and purinergic and scavenger receptors. Taken together, they highlight how the use of real-time live-cell flow cytometry is a rapid, cost-effective, reproducible, and quantifiable method to investigate P2X7 receptor function.
dc.description.peerreviewedYes
dc.languageEnglish
dc.language.isoeng
dc.publisherJournal of Visualized Experiments (JoVE)
dc.publisher.placeUnited States
dc.relation.ispartofpagefrome59313: 1
dc.relation.ispartofpagetoe59313:9
dc.relation.ispartofissue146
dc.relation.ispartofjournalJournal of Visualized Experiments (JoVE)
dc.relation.ispartofvolume146
dc.subject.fieldofresearchBiochemistry and cell biology
dc.subject.fieldofresearchPsychology
dc.subject.fieldofresearchCognitive and computational psychology
dc.subject.fieldofresearchcode3101
dc.subject.fieldofresearchcode52
dc.subject.fieldofresearchcode5204
dc.subject.keywordsScience & Technology
dc.subject.keywordsMultidisciplinary Sciences
dc.subject.keywordsScience & Technology - Other Topics
dc.subject.keywordsDevelopmental Biology
dc.subject.keywordsIssue 146
dc.titleReal-time Live-cell Flow Cytometry to Investigate Calcium Influx, Pore Formation, and Phagocytosis by P2X7 Receptors in Adult Neural Progenitor Cells
dc.typeJournal article
dc.type.descriptionC1 - Articles
dcterms.bibliographicCitationLeeson, HC; Chan-Ling, T; Lovelace, MD; Brownlie, JD; Weible, MW; Gu, BJ, Real-time Live-cell Flow Cytometry to Investigate Calcium Influx, Pore Formation, and Phagocytosis by P2X7 Receptors in Adult Neural Progenitor Cells, Journal of Visualized Experiments (JoVE), 2019, 146 (146), pp. e59313: 1-e59313:9
dcterms.licensehttp://creativecommons.org/licenses/by-nc-nd/3.0/
dc.date.updated2020-02-21T03:52:18Z
dc.description.versionVersion of Record (VoR)
gro.rights.copyright© The Author(s) 2019. This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported (CC BY-NC-ND 3.0) License, which permits unrestricted, non-commercial use, distribution and reproduction in any medium, providing that the work is properly cited.
gro.hasfulltextFull Text
gro.griffith.authorWeible, Michael W.
gro.griffith.authorBrownlie, Jeremy


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