Correlation of in vitro cell viability and cumulative singlet oxygen luminescence from protoporphyrin IX in mitochondria and plasma membrane.

SIGNIFICANCE: Photodynamic therapy (PDT) can be targeted toward different subcellular localizations, and it is proposed that different subcellular targets vary in their sensitivity to photobiological damage. Since singlet oxygen (1 O2 ) has a very short lifetime with a limited diffusion length in cellular environments, measurement of cumulative 1 O2 luminescence is the most direct approach to compare the PDT sensitivity of mitochondria and plasma membrane.

APPROACH: PDT-generated near-infrared 1 O2 luminescence at 1270 nm was measured together with cell viability for 5-aminolevulinic acid (ALA)-induced protoporphyrin IX (PpIX) and exogenous PpIX, at different incubation times. Confocal fluorescence microscopy indicated that ALA-induced PpIX (2 h) localized in the mitochondria, whereas exogenous PpIX (1 h) mainly localized to the plasma membrane. Cell viability was determined at several time points during PDT treatments using colony-forming assays, and the surviving fraction correlated well with cumulative 1 O2 luminescence counts from PpIX in mitochondria and plasmas membrane, respectively.

RESULTS: The mitochondria are more sensitive than the plasma membrane by a factor of 1.7.

CONCLUSIONS: Direct 1 O2 luminescence dosimetry's potential value for comparing the PDT sensitivity of different subcellular organelles was demonstrated. This could be useful for developing subcellular targeted novel photosensitizers to enhance PDT efficiency.