A prototype instrument is described, based on optical fiber light delivery and detection, for quantitative, non-invasive, in vivo studies of photosensitizers used in photodynamic therapy. The primary purpose of the instrument is the measurement of absolute photosensitizer uptake in tissues. This is achieved by measuring the spectrum of diffusely reflected light and identifying the characteristic absorption spectrum of the photosensitizer. This absorbance measurement is converted to equivalent absolute photosensitizer concentration by a procedure which separately determines the intrinsic optical absorption and transport scatter coefficients of the tissue. The procedure involves measuring the radial dependence of the diffusely reflected light on the tissue surface as a function of wavelength. The principles and operation of the instrument have been evaluated in tissue-simulating phantoms and in normal and tumor tissues in animal models in vivo. The outstanding problems and future development of this method, and of analogous quantitative measurements of photosensitizer fluorescence in vivo are discussed.
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