Nonlinear Behavior of the Autofluorescence Intensity on the Surface of Light-Scattering Biotissues and its Theoretical Proof.

In the up-to-date medical laser fluorescence spectroscopy (LFS) in vivo, there is a problem of quantification of fluorophores concentrations in optically-turbid biotissues by measurements of the laser induced autofluorescence flux on the surface of these tissues. One of the main problems is: whether the flux depends linearly or non-linearly on the concentration of fluorophores in tissues? The purpose of this work was both experimental and theoretical study of the character of dependencies between measured fluorescence intensities and fluorophores concentrations in optically-turbid media. In the experimental part of our study, measurements of the superficial fluorescence on phantoms at various known concentrations of fluorophores in them were carried out. As a result, experimental dependencies of registered intensities of the laser-induced autofluorescence emission were plotted against fluorophore concentrations. In the theoretical part of the study, the analytical solution for the fluorescence emission by Kokhanovsky's method based on the well-known two-flux Kubelka-Munk approach (KMA) was used. In addition, in our study the Kokhanovsky's method was modified by its association with our improved KMA, allowing us to receive exact analytical solutions for boundary intensities collected by optical probes. As a result, a set of theoretical curves describing the influence of fluorophore concentrations on the registered autofluorescence intensities was obtained, as well. Both experimental and theoretical results show a good qualitative agreement with each other. Also, these results demonstrate that the dependence of the fluorescence intensity on tissues' optical properties and on the concentration of fluorophores in light-scattering tissues can be both nonlinear and non-monotonic.