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

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.

Photophysical properties of novel fluorescein derivative and its applications for time-resolved fluorescence spectroscopy

“…These responses are almost identical. Fl-BHQ2 compound can be conveniently used to observe the responses of other slower detectors prompt to the 'color effect', as has been done recently for avalanche photodiodes used commonly in a time-resolved microscopy [1,9,10]. The advantage of using an intramolecular quenched system is a lower concentration of quencher used and independence on the viscosity of the solvent.…”

“…Already three decades ago Harris and Lytle [8] used KI to quench Rhodamine 6G. In the recent works we presented two ultra-short fluorescence standards Erythrosine B quenched by KI to about 25 ps [9] and Rose Bengal which has a lifetime of 16 ps in saturated KI water solution [10]. A reduced lifetime can be also achieved by a Foerster resonance energy transfer (FRET) mechanism [1,11].…”

USP29 is a novel non-canonical Hypoxia Inducible Factor-α activator

“…Exact time between laser excitation and photon arrival was recorded by the Time-correlated single photon counting device (TCSPC) TimeHarp260 (PicoQuant) and plotted in a histogram, thereby building up a fluorescence decay curve. An instrument response function (IRF) using erythrosine B was recorded in the same measurement conditions on an everyday basis as described previously (Szabelski et al,2009). SymPhoTime 64 software (PicoQuant) controlled all PicoQuant hardware devices and was used for analysis.…”

Multidimensional visualization of healthy and sensitized rabbit knee tissues by means of confocal microscopy

“…The instrument response function (IRF) was measured using a dye, erythrozine B. 27 A spectrometer QE65000 (Ocean Optics Inc.) was connected to the confocal scanning C1si system of the microscope to measure the integral fluorescence spectra using certain integration times during the sequential scanning of samples with a 405 nm beam of the CW diode laser. The light emitted from the tissue was collected through a 500 nm long-pass filter ("ЖC 18," LOMO, Russia) and directed to the tip of a fiber connected directly to the spectrometer.…”