Fluorescence multi-fluorophore solution

Fluorescence decays obtained for pure solutions, in the absence of fluorophore-solvent interactions, are perfectly described by mono-exponential functions (a single decay time is obtained). Hotvever, the fluorescence decay of multi-fluor-ophore solutions (usually the case of complex systems) requires the adjustment of multi-exponential functions (Figure 12.3). Note that a solution containing a single fluorophore in different environments should be considered as a mixture of fluorophores. 

Spectral imaging has been implemented in several ways on widefield or con-focal microscopes. Some of the solutions require specific instrumentation, but the method is also very generally applicable since any multi-channel fluorescence image can be considered as a series of spectral images. On a standard widefield fluorescence microscope, spectral separation of overlapping fluorophores was shown to be improved by determining and correcting for the crossover of individual fluorophores into different filter sets [15,16]. Fourier... 

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