title: The use of flow cytometry to examine calcium signalling by TRPV1 in mixed cell populations.
abstract: Flow cytometric analysis of calcium mobilisation has been in use for many years in the study of specific receptor engagement or isolated cell:cell communication. However, calcium mobilisation/signaling is key to many cell functions including apoptosis, mobility and immune responses. Here we combine multiplex surface staining of whole spleen with Indo-1 AM to visualise calcium mobilisation and examine calcium signaling in a mixed immune cell culture over time. We demonstrate responses to a TRPV1 agonist in distinct cell subtypes without the need for cell separation. Multi parameter staining alongside Indo-1 AM to demonstrate calcium mobilization allows the study of real time calcium signaling in a complex environment.
authors.affiliation: Faculty of Biology Medicine and Health, University of Manchester, UK; Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, King AbdulAziz University, Jeddah, Saudi Arabia. Electronic address: Massas@kdu.edu.sa.
authors.forename: Bakri M
authors.initials: BM
authors.lastname: Assas
authors.affiliation: Department of Biochemistry, Cancer Metabolism and Epigenetic Unit, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia.
authors.forename: Wesam H
authors.initials: WH
authors.lastname: Abdulaal
authors.affiliation: Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, King AbdulAziz University, Jeddah, Saudi Arabia.
authors.forename: Majed H
authors.initials: MH
authors.lastname: Wakid
authors.affiliation: Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, King AbdulAziz University, Jeddah, Saudi Arabia.
authors.forename: Haytham A
authors.initials: HA
authors.lastname: Zakai
authors.affiliation: Faculty of Biology Medicine and Health, University of Manchester, UK.
authors.forename: J
authors.initials: J
authors.lastname: Miyan
authors.affiliation: Faculty of Biology Medicine and Health, University of Manchester, UK.
authors.forename: J L
authors.initials: JL
authors.lastname: Pennock
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000235:genetics / Q000276:immunology; D001402:B-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D015415:Biomarkers; D002118:Calcium / Q000276:immunology / Q000378:metabolism*; D020013:Calcium Signaling; D002211:Capsaicin / Q000494:pharmacology; D003713:Dendritic Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D005434:Flow Cytometry / Q000379:methods*; D015870:Gene Expression; D007211:Indoles / Q000737:chemistry; D008264:Macrophages / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D061251:Primary Cell Culture; D013154:Spleen / Q000166:cytology / Q000276:immunology; D013194:Staining and Labeling / Q000379:methods*; D013601:T-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D050916:TRPV Cation Channels / Q000172:deficiency / Q000235:genetics* / Q000276:immunology
chemical_list: D015703:Antigens, CD; D015415:Biomarkers; D007211:Indoles; D050916:TRPV Cation Channels; C493921:TRPV1 protein, mouse; C059587:Indo-1 pentaacetoxymethyl ester; D002211:Capsaicin; D002118:Calcium
keywords: Calcium signaling; Flow cytometry; Immune phenotyping; Indo-1 AM; TRPV1