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Improved method for measuring intracellular Ca++ with fluo-3.

G T Rijkers·L B Justement·A W Griffioen·J C Cambier
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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.

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Aniline CompoundsAnimalsB-LymphocytesBenzopyransCalciumFluorescent DyesFluorometryHumansHydrogen-Ion ConcentrationLeukocytes, MononuclearXanthenesJournal ArticleResearch Support, Non-U.S. Gov't

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G T Rijkers, L B Justement, A W Griffioen, J C Cambier. 1990. Improved method for measuring intracellular Ca++ with fluo-3.. https://doi.org/10.1002/cyto.990110813

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