Photophysical study of the Ca2+ indicator Fura-2 and the K+ indicator PBFI

Viviane Van den Bergh; Katrien Meuwis; Noel Boens; Frans C. De Schryver; Michel Vincent; Jacques Gallay; Marcel Ameloot

doi:10.1117/12.182705

17 August 1994 Photophysical study of the Ca²⁺ indicator Fura-2 and the K⁺ indicator PBFI

Viviane Van den Bergh, Katrien Meuwis, Noel Boens, Frans C. De Schryver, Michel Vincent, Jacques Gallay, Marcel Ameloot

Proceedings Volume 2137, Time-Resolved Laser Spectroscopy in Biochemistry IV; (1994) https://doi.org/10.1117/12.182705
Event: OE/LASE '94, 1994, Los Angeles, CA, United States

Abstract

The fluorescent indicators Fura-2 and PBFI are widely used for the determination of intracellular concentrations of Ca²⁺ and K⁺, respectively. To investigate the complex forming reaction between Fura-2 and Ca²⁺, and between PBFI and K⁺ in the ground and excited states, steady-state and time-resolved measurements were performed. The fluorescence decay surfaces were analyzed with global compartmental analysis yielding the following values for the rate constants at room temperature in aqueous solution: (1) for Fura-2: k₀₁ equals 1.2 X 10⁹ s^-1, k₂₁ equals 1.0 X 10¹¹ M^-1x^-1, k₀₂ equals 5.5 X 10⁸s^-1, k₁₂ equals 2.2 X 10⁷s^-1 (2) for PBFI: k₀₁ equals 1.1 X 10⁹s^-1, k₂₁ equals 2.7 X 10⁸M^-1s^-1, k₀₂ equals 1.8 X 10⁹s^-1, k₁₂ equals 1.4 X 10⁹s^-1 k₀₁ and k₀₂ denote the deactivation rate constants of the free and bound forms of the indicator, respectively k₂₁ represents the bimolecular rate constant of binding of the cation by the indicator whereas k₁₂ is the rate constant of dissociation of the cation:indicator complex. For both probes the effect of the excited-state reaction can be neglected in the determination of K_d and/or the ion concentration.

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Viviane Van den Bergh, Katrien Meuwis, Noel Boens, Frans C. De Schryver, Michel Vincent, Jacques Gallay, and Marcel Ameloot "Photophysical study of the Ca²⁺ indicator Fura-2 and the K⁺ indicator PBFI", Proc. SPIE 2137, Time-Resolved Laser Spectroscopy in Biochemistry IV, (17 August 1994); https://doi.org/10.1117/12.182705

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