The ROS induced by laser illumination and its effect on cell migration

• Main Authors: Eugene Youjhen Lu, 盧宥蓁
• Other Authors: Chau-Hwang Lee
• Format: Others
• Language: zh-TW
• Published: 2014
• Online Access: http://ndltd.ncl.edu.tw/handle/66548515169039697065

碩士 === 國立陽明大學 === 生醫光電研究所 === 102 === We project a 473 nm and a 660 nm laser light pattern onto the cell adhesion surface to modulate cell migration. And we investigate the role of reactive oxygen species (ROS) in the mechanism of how blue light induces cell migration behavior. A spatial light modulator(SLM) is used to generate 473 nm and 660 nm laser pattern imaged onto the stage of a microscope. This system was created by Dr. Jian-Long, Xiao. Through this system, our group have demonstrated that 473 nm laser light pattern can be used to culture cell into arbitrary shapes of cell sheets, or push cell move forward. We also used 473 nm laser pattern as a tool to study collective migration of lung cancer cells CL1-5. Compared to other methods such as micro fluidic systems, using light to create channels to study cell migration provides a better temporal and spatial resolution. For example, we can easily change the shape of optical channels at any time. However, the mechanism of optical modulation on cell migration is still unclear. From the report of other groups, we know that ROS play an important role when cells are damaged by blue light. And ROS is also related to cell migration in respond to environmental cues. We use ROS indicator (CM-H2DCFDA) to label cellular ROS when A549 cell is illuminated by a 473 nm and 660 nm laser pattern. The result shows that the ROS level induced by the 473 nm illumination has a dose dependency on the illumination intensity, but not in the 660 nm case. We also found that cell can sense a sharp intensity gradient of 473 / 660 nm laser and migrate to the dark/ bright side. Finally, we found that the ROS has no special distribution in a single cell, which suggest that CM-H2DCFDA labeled ROS is not involved in 473 nm laser induced cell migration.