Emission Ratiometric Imaging of Intracellular Zinc: Design of a Benzoxazole Fluorescent Sensor and Its Application in Two-Photon Microscopy

Masayasu Taki, Janet L. Wolford, Thomas V. O'Halloran*

*Corresponding author for this work

Research output: Contribution to journalArticle

457 Scopus citations

Abstract

Zinc and calcium are ubiquitous intracellular metals, and while a variety of quantitative probes have been developed for measuring intracellular changes in calcium concentration, the same is not true of zinc. We describe here the design, synthesis, and properties of the benzoxazole-based, ratiometric zinc probe, Zinbo-5. This bright fluorescent reporter has a quantum yield of 0.1 in the zinc-form, exhibits a Kd for Zn2+ in the nanomolar range, and shows significant changes in both excitation and emission maxima upon zinc binding. The utility of this cell permeable probe is demonstrated in fluorescence microscopy emission ratio imaging experiments on mammalian cells. We further show that Zinbo-5 is well suited for two-photon excitation microscopy ratio imaging and can readily reveal changes in intracellular zinc concentration within optical planes of single cells. To the best of our knowledge, this is the first example of two-photon excitation microscopy applied to ratio imaging of zinc. These methods can be applied to real-time emission or excitation ratio imaging studies of zinc physiology in living cells.

Original languageEnglish (US)
Pages (from-to)712-713
Number of pages2
JournalJournal of the American Chemical Society
Volume126
Issue number3
DOIs
StatePublished - Jan 28 2004

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

Fingerprint Dive into the research topics of 'Emission Ratiometric Imaging of Intracellular Zinc: Design of a Benzoxazole Fluorescent Sensor and Its Application in Two-Photon Microscopy'. Together they form a unique fingerprint.

  • Cite this