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Intracellular Photobleaching of 5,10,15,20‐Tetrakis(m‐hydroxyphenyl) chlorin (Foscan®) Exhibits a Complex Dependence on Oxygen Level and Fluence Rate ¶

The understanding of photosensitizer photobleaching is important not only for mechanistic studies, but also for the development of monitoring techniques for clinical dosimetry in photodynamic therapy. In this study, we investigated the intracellular photobleaching of 5,10,15,20‐tetrakis(m‐hydroxyphenyl)chlorin (mTHPC, Foscan®) in the murine macrophage cell line J774A.1, using quantitative fluorescence imaging microscopy, microspectrofluorometry and microspectrophotometry. Using 652 nm laser irradiation, it was found that mTHPC exhibits oxygen‐ and fluence rate–dependent intracellular photobleaching. The kinetics showed an inverse dose–rate behavior, i.e. a reduction of fluence rate resulted in more photobleaching at comparable fluences. The effect of deoxygenation was found to be more complex, with decreased bleaching at low fluence rates and increased bleaching at higher fluence rates. The intracellular formation of reactive oxygen species was measured using 2′,7′‐dichlorodihydrofluorescein diacetate. The results are analyzed in terms of competitive Type‐I and Type‐II mechanisms.