Time-resolved fluorescence spectroscopy lifetime analysis

This interpretation was confirmed by time-resolved fluorescence spectroscopy using the single photon timing technique. The pentad as a 1 X 10 M solution in chloroform was excited at 590 nm, and emission decay curves were recorded at 14 wavelengths. All 14 decays were then fit simultaneously to four exponential functions (x = 1.12) using a global analysis technique. The results were used to construct the decay associated spectrum shown in Figure 16. The two major components of the decay had lifetimes of 0.039 and 1.2 ns. (The two minor components represent impur-... 

Wang X. F., Periasamy A., Wodnicki P., Gordon G. W. and Herman B. (1996) Time-Resolved Fluorescence Lifetime Imaging Microscopy Instrumentation and Biomedical Applications, in Wang X. F. and Herman B. (Eds), Fluorescence Imaging Spectroscopy and Microscopy, Chemical Analysis Series, Vol. 137, John Wiley ... 

For the determination of the dissociation constant in the excited state, several methods have been used the Forster cycle,(109 m) the fluorescence titration curve/113 the triplet-triplet absorbance titration curve,014 but all involve the assumption that the acid-base equilibrium may be established during the lifetime of the excited state, which is by no means a common occurrence. A dynamic analysis using nanosecond or picosecond time-resolved spectroscopy is therefore often needed to obtain the correct pK a values.1(n5)... 

A general discussion of the use of least-squares fitting in fluorescence measurements may be found in (28). The global analysis of fluorescence data is discussed in (29). Commercially available time-resolved fluorimeters are typically sold with data analysis software included. Available stand-alone packages include the Globals Unlimited suite, which is capable of analysing both time- and frequency-domain data, stopped-flow kinetics, etc. The Center for Fluorescence Spectroscopy at the University of Maryland (USA) also offers software for frequency- and time-domain fluorescence lifetime analysis. 

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