title: Improved method for measuring intracellular Ca++ with fluo-3.
abstract: The accuracy of flow cytometric measurement of intracellular calcium with fluo-3 is compromised by variation in basal fluorescence intensity due to heterogeneity in dye uptake or compartmentalization. We have loaded cells simultaneously with fluo-3 and SNARF-1. When SNARF-1 fluorescence is collected at approximately 600 nm, its intensity does not change upon cell activation. Furthermore, fluo-3 and SNARF-1 fluorescence signals exhibit a linear relationship. The ratio of fluo-3 to SNARF-1 eliminates a significant proportion of variation in fluorescence intensity caused by variation in fluo-3 uptake and thus can be used as a sensitive parameter for measuring changes in [Ca2+]i.
authors: Department of Immunology, University Hospital for Children and Youth, Het Wilhelmina Kinderziekenhuis, Utrecht, The Netherlands.
authors: G T
authors: GT
authors: Rijkers
authors: L B
authors: LB
authors: Justement
authors: A W
authors: AW
authors: Griffioen
authors: J C
authors: JC
authors: Cambier
mesh_terms: D000814:Aniline Compounds; D000818:Animals; D001402:B-Lymphocytes / Q000737:chemistry; D001578:Benzopyrans; D002118:Calcium / Q000032:analysis* / Q000378:metabolism; D005456:Fluorescent Dyes; D005470:Fluorometry / Q000379:methods; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007963:Leukocytes, Mononuclear / Q000737:chemistry; D014966:Xanthenes
chemical_list: D000814:Aniline Compounds; D001578:Benzopyrans; D005456:Fluorescent Dyes; C063917:SNARF dye; D014966:Xanthenes; C059715:Fluo-3; D002118:Calcium