Grynkiewicz G, Poenie M, Tsien R Y
J Biol Chem. 1985 Mar 25;260(6):3440-50.
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.
为了对胞质游离钙离子的生理作用进行生化研究，已经合成了一类新型的高荧光指示剂。这些化合物将一个八配位的四羧酸盐螯合位点与芪发色团结合在一起。将芪的烯键并入杂环可提高荧光团的量子效率和光化学稳定性。与广泛使用的前身“quin2”相比，新染料的荧光亮度提高了30倍，钙离子结合时不仅强度发生变化，波长也有重大改变，对钙离子的亲和力略低，激发波长略长，对钙离子相对于其他二价阳离子的选择性有显著提高。这些特性，尤其是对钙离子的波长敏感性，应使这些染料成为许多细胞内应用的首选荧光指示剂，特别是在单细胞、贴壁细胞层或大块组织中。
