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Photophysics of the Fluorescent pH Indicator BCECF (CROSBI ID 125097)

Prilog u časopisu | izvorni znanstveni rad | međunarodna recenzija

Boens, Noël ; Qin, Wenwu ; Basarić, Nikola ; Orte, Angel ; Talavera, Eva M. ; Alvarez-Pez, Jose M. Photophysics of the Fluorescent pH Indicator BCECF // The journal of physical chemistry. A, 110 (2006), 30; 9334-9343-x

Podaci o odgovornosti

Boens, Noël ; Qin, Wenwu ; Basarić, Nikola ; Orte, Angel ; Talavera, Eva M. ; Alvarez-Pez, Jose M.

engleski

Photophysics of the Fluorescent pH Indicator BCECF

The photophysical behavior of BCECF [2', 7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein] − currently the most widely used fluorescent pH indicator for near-neutral intracellular pH measurements – has been explored by using absorption and steady-state and time-resolved fluorescence measurements. The influence of ionic strength as well as total buffer concentration on the absorbance and steady-state fluorescence has been investigated. The apparent acidity constant of the pH indicator determined by absorbance and fluorescence titration is dependent on the added buffer and salt concentrations. A semi-empirical model is proposed to rationalize the variations in the apparent pKa values. The excited-state proton exchange of BCECF at physiological pH becomes reversible upon addition of phosphate buffer inducing a pH dependent change of the fluorescence decay times. Fluorescence decay traces collected as a function of total buffer concentration and pH were analyzed by global compartmental analysis yielding the following values of the rate constants describing excited-state dynamics of BCECF: k01 = 3.4 x 108 s− 1, k02 = 2.6 x 108 s− 1, k21 = 1 x 106 M− 1 s− 1, = 1.4 x 108 M− 1 s− 1, and = 4.3 x 107 M− 1 s− 1.

absorption; BCECF; fluorescence; global analysis; photophysics; pH

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Podaci o izdanju

110 (30)

2006.

9334-9343-x

objavljeno

1089-5639

Povezanost rada

Kemija

Indeksiranost