Collisional Quenching of Erythrosine B as a Potential Reference Dye for Impulse Response Function Evaluation

Authors: Szabelski, Mariusz¹; Ilijev, Douglas²; Sarkar, Pabak²; Luchowski, Rafal²; Gryczynski, Zygmunt³; Kapusta, Peter⁴; Erdmann, Rainer⁴; Gryczynski, Ignacy³

Source: Applied Spectroscopy, Volume 63, Issue 3, Pages 68A-86A and 261-380 (March 2009) , pp. 363-368(6)

Publisher: Society for Applied Spectroscopy

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Abstract:

We studied the collisional quenching of the erythrosine B fluorophore by potassium iodide. The quenching follows a Stern-Volmer dependence up to the highest quencher concentration. The lifetime of erythrosine B decreases to 24 ps in 5.02 M of potassium iodide. The quantum yield of erythrosine B in the presence of 5.02 M KI is 0.0035. The relatively high brightness makes this compound attractive as an ultrashort reference in time-resolved measurements. In both frequency- and time-domain fluorescence techniques, there is a need for lifetime standards with extremely short decay times. Mimicking the instantaneous scattering at longer wavelengths allows color-effect-free measurements in the emission region. Another motivation is the problem of obtaining the impulse response function in the case of two-photon excitation. Time-resolved microscopy also benefits from fast-decaying dyes because the impulse response function can be evaluated at the emission wavelength of the investigated specimen without changing filters. We demonstrated that impulse response functions for commonly used detectors are practically the same for scattering as for quenched erythrosine B emission. We also analyzed a complex fluorescence decay using both elastic scattering and quenched erythrosine B emission as a response function.

Keywords: ERYTHROSINE B; IMPULSE RESPONSE FUNCTION; LIFETIME; COLLISIONAL QUENCHING

Document Type: Research article

DOI: 10.1366/000370209787598979

Affiliations: 1: Center for Commercialization of Fluorescence Technologies, Department of Molecular Biology and Immunology, UNT HSC, Fort Worth, Texas; Institute of Experimental Physics, University of Gdansk, Wita Stwosza 57, 80-952 Gdansk, Poland 2: Center for Commercialization of Fluorescence Technologies, Department of Molecular Biology and Immunology, UNT HSC, Fort Worth, Texas 3: Center for Commercialization of Fluorescence Technologies, Department of Molecular Biology and Immunology, UNT HSC, Fort Worth, Texas; Department of Cell Biology and Genetics, UNT HSC, Fort Worth, Texas 4: PicoQuant GmbH, Berlin, Germany

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