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. 1985 Mar 25;260(6):3440-50.

A new generation of Ca2+ indicators with greatly improved fluorescence properties

G Grynkiewicz, M Poenie, R Y Tsien
  • PMID: 3838314
Free article

A new generation of Ca2+ indicators with greatly improved fluorescence properties

G Grynkiewicz et al. J Biol Chem. .
Free article

Abstract

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.

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