Recovery of turbidity free fluorescence from measured fluorescence: an experimental approach.

Fluorescence from fluorophores embedded in a turbid medium like biological tissue gets strongly modulated by the wavelength dependent absorption and scattering properties of tissue. This makes it extremely difficult to extract valuable biochemical information from tissue which is present in the intrinsic line shape and intensity of fluorescence from tissue fluorophores. We present an experimental approach to remove the distorting effect of scattering and absorption on intrinsic fluorescence of fluorophores embedded in a turbid medium like tissue. The method is based on simultaneous measurement of polarized fluorescence and polarized elastic scattering spectra from a turbid medium. The polarized fluorescence normalized by the polarized elastic scattering spectra (in the wavelength range of fluorescence emission) was found to be free from the distorting effect of absorption and scattering properties of the medium. The applicability range of this technique to recover intensity and line shape information of intrinsic fluorescence has been investigated by carrying out studies on a variety of tissue phantoms having different absorption and scattering properties. The results obtained show that this technique can be used to recover intrinsic line shape and intensity information of fluorescence from fluorophores embedded in a scattering medium for the range of optical transport parameters typically found in biological tissue.