Abstract
High-precision resolution for multi-exponential fluorescence decay is obtained by completely automated multifrequency phase fluorometry, using the harmonic content of the pulse train of a stabilised, mode-locked, cavity-dumped dye laser. The principle of this method has been described previously (Lackowicz et al. 1986). By stabilising the pump laser, the frequency stability of the cross correlation product was considerably enhanced. The use of a flat-top band-pass filter enables fast measurement and ensures high phase stability. Phase shifts with a standard deviation of 0.1 degree are measured at the harmonics of the basic frequency (400 kHz) with a digital counter. All instruments, including monochromator and cell exchanger, are fully computer-controlled. The instrument has been used successfully to elucidate the complex fluorescence decay of proteins, antibiotics and some inorganic Cu complexes.
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