A Comparison of Fluorescence Inner-Filter Effects for Different Cell Configurations

Abstract

In conventional fluorescence spectroscopy, fluorescence intensity at high fluorophore concentration is oRen not proportional to fluorophore concentration, owing to primary and secondary absorption (inner-filter effects). In this paper, fluorescence calibration curves for anthracene solutions were obtained using a conventional right angle cell, a frontal reflection cell, a short pass cell, and a total internal reflection fluorescence (TIRF) cell for comparing the observed primary inner-filter effects. Measurements were also made of a two-component mixture using the nonfluorescent 94trofluorene with anthracene for comparing primary and secondary inner-filter effects. A conventional right angle cell exhibited the widest linear dynamic range and lowest detectable anthracene concentration, whereas the TIRF cell provided the best linearity at high concentrations. The TIRF cell was determined to have significant potential for quantitative analysis of hghly concentrated and/or turbid solutions.

Total synchronous fluorescence scan spectra of petroleum products

“…Excitation source was a 100 W Xenon lamp. Right-angle geometry was used for measurement because for fluorophores at higher concentrations it gives larger linear dynamic range [22], lower detection limit [22] and larger shift in synchronous fluorescence maximum [6,7] as compared to the front-face or 45°-angle sample geometry. The dimension of the cell used was 1 cm 2 .…”

Inner Filter Effect in the Fluorescence Emission Spectra of Room Temperature Ionic Liquids with-β-Carotene

“…The all-trans β-carotene in standard solvent, for example hexane, fluorescence quantum yield of electronic state S 2 is 2 x 10 -4 (Sherve et al, 1991;Andersson, 1992) and of S 1 state is 4 x 10 -6 (Wasielewski, 1986(Wasielewski, , 1989 Inner filter effects are often responsible for distorted emission spectra and nonlinear calibration curves between fluorescence intensity and fluorophore concentration. In a case of concentrated samples, in order to minimize or correct for IFE either instrumental or mathematical corrections can be done (Kao et al, 1998). Instrumental corrections are based on the observation that fluorescence intensity and fluorophore concentration are linear at low absorbance.…”

“…The serious problem of quantitative analysis is dominant in the samples with optical density higher than 0.05 (10 mm cell) detected in the right angle cell geometry, because of the two kinds of inner filter effects IFEs (Lakowicz, 2006;Li & Hu, 2007;Kao et al, 1998). The primary inner filter effect (PIFE) -defined as the decrease in the intensity of the excitation beam at the point of observation because of the chromophore optical absorption in the excitation region.…”

Luminescent method for determining low concentrations of a substance in optically dense media

“…The geometry was varied by using different relative orientations of the exciting and detected fluxes: at a right angle, in-line, and frontal [1,8,9]. The function f(x) was varied by modification of the system for collecting the radiation and the thickness d of the cuvet.…”