Time resolved fluorescence overview

Volume 4 is intended to summarize the principles required for these biomedical applications of time-resolved fluorescence spectroscopy. For this reason, many of the chapters describe the development of red/NIR probes and the mechanisms by which analytes interact with the probes and produce spectral changes. Other chapters describe the unique opportunities of red/NIR fluorescence and the types of instruments suitable for such measurements. Also included is a description of the principles of chemical sensing based on lifetimes, and an overview of the ever-important topic of immunoassays. 

Although the emission from polymers with pendant aromatic groups is complex and there remains controversy concerning the specific kinetic models required to describe their fluorescence decay behaviour, the results presented above illustrate the usefulness of time-resolved fluorescence surfaces for obtaining an overview of the energy relaxation process in polymeric systems. 

The application of time-resolved fluorescence spectroscopy to studies of excimer formation and energy transfer in PACE and P2NMA provides an overview of the emitting species present and the dynamics of energy relaxation in these polymers. The results of fluorescence decay analyses suggest that kinetic models which have been proposed to explain monomer/excimer kinetics may require further refinanent. 

An analysis of the time-resolved fluorescence in excimer-forming polymer systems can also prove informative in elucidating the conformations of macromo-lecular chains in solution these data are particularly relevant in terms of the structures adopted by polyelectrolytes in aqueous media. In this latter context, it is useful at this stage to briefly overview the development of the kinetic schemes to describe excited state interactions in fluorescent polymer solutions. 

This overview on applications of time-resolved fluorescence techniques can only give a few examples of the many uses in biochemistry, biology, and clinical chemistry. It is not difficult to predict that applications... 

It may not be evident to the reader, but 1 have attempted to keep this chapter as brief as possible. There have been numerous developments in the technology since 1983, and I felt these needed to be mentioned for acomplete overview of the technology for time-resolved fluorescence. Numer< ous additional details could have, and perhtqM should hav e, been described. It is hoped that this chapter provides the basics for understanding the extensive literature on doie-resolved fluorescence. 

In the previous chapter we presented an overview of protein fluorescence. We described the spectral properties of the aromatic amino acids and how these properties depend on protein structure. We now extend this discussion to include time-resolved measurements of intrinsic protein flu( scence. Prior to 1983, most measurements of time-resolved fluorescence were performed using TCSPC. The instruments employed for these measurements typically used a flashlamp etcitation source and a standard dynode-chain-type PMT. Such instruments provided instrument response functions with a half-width near 2 ns, which is comparable to thedecay time of most proteins. The limited repetition rate of the flashlamps, near 20 kHz. resulted in data of modest statistical accuracy, unless the acquisition times were excessively long. Given the complexity of protein intensity and anisotropy decays, and the inherent difficulty of resolving multiexponential processes. ii was difficult to obUun definitive information on the decay kinetics of proteins. 

See also Fluorescence Overview Multidimensional Fluorescence Spectrometry Time-Resolved Fluorescence Derivatization Quantitative Analysis Environmental Applications Food Applications. 

See also Chemiluminescence Overview. Fluorescence Time-Resolved Fluorescence. Immunoassays, Techniques Enzyme Immunoassays. Phosphorescence Room-Temperature. Sequential Injection Analysis. 

Table 1 gives an overview of the various hardware configurations for wide-field time-resolved fluorescence microscopy (TRFM). 

In this second edition of Principles of Fluorescence Spectroscopy, I have attempted to maintain the emj asis on basics, while updating the examples to include more recent results from the literature. Iliere is a new chapter providing an overview of extrinsic fluoroj ores. The discussion of time-resolved measurements has been ex-... 

The present article gives a short overview on some methods of fluorescence spectroscopy and microscopy with some emphasis on time-resolving techniques. Examples for application are concentrated on the well known mitochondrial marker... 

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