Far-red fluorescence probe for monitoring singlet oxygen during photodynamic therapy

Article abstract of DOI:10.1021/ja504279r

Singlet oxygen (¹O₂), molecular oxygen in the lowest excited state, has a critical role in the cell-killing mechanism of photodynamic therapy (PDT). Although ¹O₂ phosphorescence measurement has been mainly used to monitor ¹O₂ formation during PDT, its intensity is far insufficient to obtain two-dimensional images of intracellular ¹O₂ with the subcellular spatial resolution using the currently available near-IR detector. Here, we propose a new far-red fluorescence probe of ¹O₂, namely, Si-DMA, composed of silicon-containing rhodamine and anthracene moieties as a chromophore and a ¹O₂ reactive site, respectively. In the presence of ¹O₂, fluorescence of Si-DMA increases 17 times due to endoperoxide formation at the anthracene moiety. With the advantage of negligible self-oxidation by photoirradiation (θ_δ < 0.02) and selective mitochondrial localization, Si-DMA is particularly suitable for imaging ¹O₂ during PDT. Among three different intracellular photosensitizers (Sens), Si-DMA could selectively detect the ¹O₂ that is generated by 5-aminolevulinic acid-derived protoporphyrin IX, colocalized with Si-DMA in mitochondria. On the other hand, mitochondria-targeted KillerRed and lysosomal porphyrins could not induce fluorescence change of Si-DMA. This surprising selectivity of Si-DMA response depending on the Sens localization and photosensitization mechanism is caused by a limited intracellular ¹O₂ diffusion distance (～300 nm) and negligible generation of ¹O₂ by type-I Sens, respectively. For the first time, we successfully visualized ¹O ₂ generated during PDT with a spatial resolution of a single mitochondrial tubule.