Adopting laser scanning confocal microscopy in dynamic observation of photodamage of subcellular sites / 中华物理医学与康复杂志

ABSTRACT

Objective To dynamically observe photodamage of subcellular sites by use of laser scanning confocal microscopy (LSCM). Methods The samples were divided into four groups. Murine lung endothelial cells were subcultured and incubated with HMME for 24 hours. Then the cells were stained with rhodamine-123 for demonstration of mitochondria. LSCM was applied and organelle-cell fluorescence intensity ratio analysis was adopted to study the intracellular distribution of HMME. Then dynamic fluorescence images sequence of rhodamine-123 was collected. Results Rhodamine-123′s fluorescence images of cell sample with HMME was changed gradually during irradiation the typical characteristic of mitochondria disappeared gradually with decreasing fluorescence intensity. The fluorescence of rhodamine-123 was diffused and distributed in nuclear, while rhodamine-123′s fluorescence images of cell sample without HMME was not changed. Conclusion Mitochondria and nucleus are photodamage sites by HMME-PDT; LSCM can be applied in dynamic observation of photodamage of subcelluar sites.