Principles of Fluorescence Spectroscopy

PRINCIPLES OF FLUORESCENCE SPECTROSCOPY 275
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PROBLEMS
P7.1. Estimation of the Spectral Relaxation Time: Figure 7.56
shows time-dependent intensity decays of TNS bound to
Figure 7.56. Fluorescence impulse response functions of TNS-labeled
egg lecithin vesicles. Time-dependent intensities are shown for 390,
435, and 530 nm. From [162].
egg lecithin vesicles. 162 The wavelengths of 390, 435,
and 530 nm are on the blue, center, and red regions of
the emission spectrum. Use the data in Figure 7.56 to
calculate the spectral relaxation time for the TNSlabeled
residue. Assume that the emission at 390 nm is
dominated by the initially excited states (F) and that the
emission at 435 nm represents the TNS, unaffected by
relaxation.
P.7.2. Interpretation of Wavelength-Dependent Lifetimes: TNS
was dissolved in various solvents or bound to vesicles of
dialeoyl-L-α-phosphatidylcholine (DOPC). Apparent
phase and modulation lifetimes were measured across
the emission spectra of these samples. Explain these
data in Figure 7.57.
Figure 7.57. Fluorescence lifetimes and spectra of TNS, dissolved in various solvents and TNS bound to DOPC vesicles. Apparent phase shift (")
and modulation (!) lifetimes were measured at 30 MHz. Normalized emission spectra are shown for TNS in glycerol and bound in DOPC vesicles
(solid), and for TNS in dioxane at 25°C (dashed).