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A New Generation of Ca2+ Indicatorsw ith Greatly Improved Fluorescence Properties*

Grzegorz Grynkiewicz$, Martin Poenie, and Roger Y. TsienB
From the Department of Physiology-Anatomy, Uniuersitoyf California, Berkeley, California9 4720
A new family of highly fluorescent indicators has been synthesized forbiochemical studies of the physiological role of cytosolic free Ca2+.The compounds combine an 8-coordinate tetracarboxylate chelating sitewith stilbene chromophores. Incorporation of the ethylenic linkage ofthe stilbene into a heterocyclic ring enhances the quantum efficiencyand photochemical stability of the fluorophore. Compared to theirwidely used predecessor, ¡°quina,¡± the new dyes offer up to 30-foldbrighter fluorescence, major changes in wavelength not just intensityupon Ca2+ binding, slightly lower affinities for Ca2+, slightly longerwavelengths of excitation, and considerably improved selectivity forCa2+ over other divalent cations. These properties, particularly thewavelength sensitivity to Ca2+, should make these dyes the preferredfluorescent indicators for many intracellular applications, especiallyin single cells, adherent cell layers, or bulk tissues.

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http://d.namipan.com/d/895bf1094d3a3db93b3051fb6b519b0a8b6c40aef5a91500

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