Knowledge of the fluence distribution in biological tissue is essential for applications of lasers and light in medicine. A method using a photoactivable fluorophore as a chemical actinometer is presented to investigate the fluence (J/cm 2 ) distribution in tissue-simulating phantoms. Such a chemical actinometer provides high spatial resolution( <=20 µ m) while minimizing the disturbance of the fluence distribution. The actinometer substance, nonfluorescent in its native state, is incorporated into an acrylamide gel. Upon absorption of 351 nm radiation ( \lambdaact ), the actinometer substance becomes a fluorophore, which is excited at \lambdaex<=485 nm. Thus the spatial distribution of the emitted fluorescence ( \lambdaex>=515 nm) in the actinometer represents the fluence distribution of the activating radiation. Using histological techniques, 20 µ m sections are cut from gel-like optical phantoms containing the actinometric substance. The fluorescence intensity in the section is recordee under a standard fluorescence microscope equipped with a sensitive video camera. To simulate different biological tissues, the scattering and absorption properties of teh gel phantoms are varied over a wide range. The experimentally obtained fluence distributions are compared with theorectical models of light distribution in turbid media.
L. Lilge, T. J. Flotte, I. E. Kochevar, S. L. Jacques, F. Hillenkamp, "Photoactivatable fluorophores for the measurement of fluence in turbid media," Photochem. Photobiol.,58, 37-44 (1993).