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J. Biol. Chem., Vol. 260, Issue 6, 3440-3450, Mar, 1985
G Grynkiewicz, M Poenie and RY Tsien
A new family of highly fluorescent indicators has been synthesized for
biochemical studies of the physiological role of cytosolic free Ca2+. The
compounds combine an 8-coordinate tetracarboxylate chelating site with
stilbene chromophores. Incorporation of the ethylenic linkage of the
stilbene into a heterocyclic ring enhances the quantum efficiency and
photochemical stability of the fluorophore. Compared to their widely used
predecessor, "quin2", the new dyes offer up to 30-fold brighter
fluorescence, major changes in wavelength not just intensity upon Ca2+
binding, slightly lower affinities for Ca2+, slightly longer wavelengths of
excitation, and considerably improved selectivity for Ca2+ over other
divalent cations. These properties, particularly the wavelength sensitivity
to Ca2+, should make these dyes the preferred fluorescent indicators for
many intracellular applications, especially in single cells, adherent cell
layers, or bulk tissues.
A new generation of Ca2+ indicators with greatly improved fluorescence properties
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