Quantitative optical biopsy of liver tissue ex vivo

The intensity of the intrinsic autofluorescence of the reduced form of nicotineamide adenine dinucleotid (NADH) of biological tissue depends on the local, cellular concentration of this coenzyme. It plays a dominant role in the Krebs cycle and, therefore, serves as an indicator for the vitality of the observed cells. Due to the individually and locally varying boundary conditions and optical tissue properties, which are scattering coefficients, absorption coefficients and an anisotropy factor g, the fluorescence signal needs to be rescaled. Rescaling methods use for instance the Kubelka-Munk theory or the photon migration theory. Our rescaling method is partly based on measurements and partly theoretically derived. By combining four methods, i.e., laser-induced fluorescence (LIF) of the time-resolved signal, biochemical concentration measurements. Monte Carlo simulations with typical optical parameters and microscopic investigations, we demonstrate that simultaneous detection of the fluorescence and backscattering signal can easily and accurately provide rescaled, quantitative values for the NADH concentrations.