Biophotonics Laboratory, National Institute of Lasers and Optronics, Islamabad, 45650, Pakistan.
Lasers Med Sci. 2010 Jul;25(4):545-50. doi: 10.1007/s10103-010-0760-6. Epub 2010 Feb 20.
Experiments were carried out on cancerous HeLa cells and blood serum using a double integrating sphere and a He-Ne laser to investigate the optical properties and cellular effects due to photodynamic therapy (PDT). In the first experiment, HeLa cells were exposed to Photofrin at concentrations of 0, 10, 20, 30, 50 and 112.4 microg/ml at an irradiance of 0.2 W/cm(2) using diode laser light. Using a confocal microscope, cell debris and morphological changes in HeLa cells were recorded at different Photofrin concentrations. The results showed cell debris in HeLa cells at the highest concentration of Photofrin. In a second experiment, photobleaching was observed in HeLa cells in the presence of various concentrations of 5-aminolaevulinic acid ranging from 0-50 microg/ml. There was progressive degradation of the 635 nm peak during continuous laser irradiation at an irradiance of 0.2 W/cm(2). We conclude that cells demonstrating high initial fluorescence undergo bleaching at a faster rate than those with lower fluorescence. Finally in a third experiment, cancerous and noncancerous blood serum was irradiated at an irradiance of 0.1 W/cm(2) using a He-Ne laser in conjunction with a double integrating sphere system. Forward and back scattering of normal and malignant serum showed an exponential decrease in fluorescence amplitude. The results indicate that there is notable amplitude difference between malignant and normal blood serum with malignant blood serum showing decreased scattering. These results have important implications for photodiagnosis and photodynamic therapy.
采用双积分球和氦氖激光器对癌细胞和血清进行了实验,以研究光动力疗法(PDT)引起的光学特性和细胞效应。在第一个实验中,HeLa 细胞在辐照度为 0.2 W/cm²的二极管激光照射下,用不同浓度的 Photofrin(0、10、20、30、50 和 112.4 µg/ml)处理。通过共聚焦显微镜,在不同的 Photofrin 浓度下记录了 HeLa 细胞的细胞碎片和形态变化。结果表明,在 Photofrin 浓度最高的情况下,HeLa 细胞中有细胞碎片。在第二个实验中,观察到在不同浓度的 5-氨基酮戊酸(0-50 µg/ml)存在下,HeLa 细胞发生光漂白。在辐照度为 0.2 W/cm²的连续激光照射下,635nm 峰逐渐降解。我们得出结论,初始荧光强度较高的细胞比荧光强度较低的细胞漂白速度更快。最后,在第三个实验中,采用氦氖激光器和双积分球系统,以辐照度为 0.1 W/cm²照射癌性和非癌性血清。正常和恶性血清的前向和后向散射显示荧光强度呈指数下降。结果表明,恶性和正常血清之间存在显著的振幅差异,恶性血清的散射减少。这些结果对光诊断和光动力疗法具有重要意义。