The F#246;rster radius were calculated from absorption spectrum of F与# 246；从吸收光谱计算了福斯特半径.docVIP

Experimental Spectroscopic Measurements The fluorescence experiments were performed on a SPEX fluorolog 112 instrument (SPEX Industries, Metuchen, NJ, USA). The spectral bandwidths were for the excitation monochromator 5.6 nm, and for the emission monochromator 2.7 nm. All fluorescence spectra were corrected. A GBC 912 (GBC Scientific Equipment Pty, Ltd, Australia) was used for absorption measurements. Fluorescence lifetimes were determined using a PRA3000 instrument (PRA Inc., London, Ontario, Canada) equipped with Glan-Thompson polarisers. Light emitting diodes, NanoLED-01 and NanoLED-04 (IBH, Glasgow, Scotland, UK) were used for excitation. Excitation and emission wavelengths were selected by interference filters (Omega/Saven, Sweden) and long-pass filters (Schott Germany). The F?rster radii were calculated from molar absorptivity, of the acceptor and corrected fluorescence spectrum, of the donor according to1 (1) where (2) and (3) Here, ?2 denotes the average squared orientational part of a dipole-dipole interaction, ?d is the fluorescence quantum yield of the donor, n is the refractive index and NA is the Avogadro constant. In calculating the F?rster radius (), ?2 = 2/3 was chosen as a reference state. Refractive indices were measured and found to be n = 1.36 and 1.45 for ethanol and chloroform, respectively. For absorption measurements, the maximum absorbance was always about 0.6 while it for fluorescence measurements was less than 0.08. The distance, R, between donor and acceptor in the cassettes was estimated by using standard bond lengths. Calculated energy transfer efficiency based on F?rster′s theory, Ecalc, was determined according to2 (4) where (5) and (6) A detailed description of ?2 and notations used is given elsewhere.3 In eqs. 4 and 5, ? denotes the fluorescence lifetime. The experimental energy transfer efficiency, Eexpt, was determined from steady-state fluorescence and absorption measurements as foll