Light dose and time dependency of photodynamic cell membrane damage

We have investigated the light dose and time dependency of photodynamic cell membrane damage using electrophysiological methods. This study controls the level of cell membrane damage by precisely administration of the light dose. The photosensitizer used was 5',5"-bis(aminomethyl)-2,2':5',2"-terthiophene dihydrochloride (BAT). A confocal laser scanning microscope was used to provide rapid light activation (< 1 s) and the subsequent membrane damage was monitored using standard patch clamp techniques. In the presence of 49 microM BAT, light levels less than 0.94 J/cm2 led to a reversible depolarization (approximately 20 mV) and reduction of resistance (approximately 10%) within 3 s of illumination. Higher intensities of illumination (> 1.57 J/cm2) caused a complete and irreversible loss of membrane potential and cell membrane resistance within 8 s illumination. The threshold dose of light required to induce cell death by illumination in the presence of BAT was increased in the presence of the antioxidant Trolox-C.