Effects of inaccurate reference lifetimes on interpreting frequency-domain fluorescence data

Abstract

The effects of assigning inaccurate reference lifetimes in lifetime determinations are predicted theoretically by using standard equations. This theory leads to a method to remove reference error effects using common least-squares software. This method cannot, however, be used to deconvolute data collected with isochronal references. Uncorrected data can always be exactly solved with models containing one more degree of freedom than the true model. Monoexponential decays are fit by double-exponential decays or excited-state processes. Unimodal distributed decays often appear as discrete, double-exponential decays.

Fluorescence biosensing strategy based on energy transfer between fluorescently labeled receptors and a metallic surface

“…This was calculated by transforming the changes in the phase angle of Fig. 4b to conversion assuming a model with two decaying components (Litwiler et al, 1990). The two decaying components were assumed to have lifetimes of 3.5 and 0.7 ns corresponding respectively to the fluorescence of the antibody in the bulk solution and the antibody bound to the surface as obtained with frequency domain measurements (vide supra).…”

Determination of the transduction mechanism for optical sensors based on Rhodamine 6G-impregnated perfluorosulfonate films using steady-state and frequency-domain fluorescence

“…RESULTS AND DISCUSSION Characterization of the Instrument. Accurate fluorescence lifetime work requires the use of well-characterized reference lifetime standards (21). For conventional liquid-phase studies, the common practice is to use dilute solutions of fluorophores (with similar spectral contours to the system under study) that decay with a single lifetime (21).…”

Spectrally Resolved Frequency Domain Analysis of Multi-Fluorophore Systems Undergoing Energy Transfer

“…This data will be presented using three graphical formats: (1) a plot of apparent lifetime versus wavelength; (2) a s-s plot of (s m, s / ); and (3) an AB plot. 14-17, à The apparent lifetimes are computed from m(k) and /(k): 8,11,[18][19][20][21] s / ðkÞ ¼ 1…”

“…21 Related treatments can be found elsewhere in the literature. 11,[18][19][20][22][23][24][25] The linear transforms N (Eq. 8) and D (Eq.…”

“…9) have in the past been reported using different symbols and/or expressions. For instance, the paired linear transforms S and G, 19,20,24 and P and Q (Lakowicz and Balter, 1982a) have been previously utilized. When the paired transforms are compared relative to each other, it is recognized that the N, S, and P transforms are equivalent.…”