We present a method for developing fluorescence based diagnostic algorithms for pathologic conditions in human tissue which can be applied in real time in vivo. The method of algorithm development is stepwise. Fluorescence excitation-emission matrices are first used to identify excitation wavelength regions optimal for the differentiation of normal and pathologic tissues. At these excitation wavelengths, the morphologic and molecular basis of tissue fluorescence is determined using a combination of fluorescence and light microscopy and studies of individual biomolecules. The optical properties of these components are then measured. A model of light propagation in tissue is used to describe tissue fluorescence spectra in terms of these optical properties and the physicochemical composition of tissue. The parameters of this model are then used to develop statistically optimized diagnostic algorithms for the presence of pathology. Finally, in vivo trials of these techniques are undertaken to begin to assess their role in clinical medicine.
© 1991 Optical Society of AmericaPDF Article