Dynamics of photoinduced cell plasma membrane injury.
Dynamics of photoinduced cell plasma membrane injury.
复制标题
光诱导细胞质膜损伤的动力学。
DOI：
10.1016/s0006-3495(95)80402-9
复制
发表时间：
1995
影响因子：
3.4
通讯作者：
Yarmush,ML
中科院分区：
文献类型：
--
作者：
Thorpe,WP;Toner,M;Ezzell,RM;Tompkins,RG;Yarmush,ML
我们已经开发了一个视频显微镜系统，设计用于实时测量的单细胞损伤在光解过程中定义良好的物理化学和生物物理条件下。体外培养的黑素瘤细胞用光敏剂（PS）、二氢锡e6（SnCe 6）或免疫缀合物（与抗ICAM单克隆抗体缀合的SnCe 6）处理，并用630 nm波长的10 mW He/Ne激光照射。使用活体染料钙黄绿素-AM评估细胞膜完整性。在激光功率密度和PS浓度变化的实验中，确定了细胞破裂之前的时间滞后与估计的单态氧到细胞表面的通量成反比。裂解事件的显微镜检查表明，光诱导裂解是由质膜的点破裂引起的。这种显微镜系统的在线性质提供了一个机会，以监测细胞损伤过程的动态，并获得洞察的机制，管理光解细胞损伤过程。
We have developed a video microscopy system designed for real-time measurement of single cell damage during photolysis under well defined physicochemical and photophysical conditions. Melanoma cells cultured in vitro were treated with the photosensitizer (PS), tin chlorin e6 (SnCe6) or immunoconjugate (SnCe6 conjugated to a anti-ICAM monoclonal antibody), and illuminated with a 10 mW He/Ne laser at a 630 nm wavelength. Cell membrane integrity was assessed using the vital dye calcein-AM. In experiments in which the laser power density and PS concentration were varied, it was determined that the time lag before cell rupture was inversely proportional to the estimated singlet oxygen flux to the cell surface. Microscopic examination of the lytic event indicated that photo-induced lysis was caused by a point rupture of the plasma membrane. The on-line nature of this microscopy system offers an opportunity to monitor the dynamics of the cell damage process and to gain insights into the mechanism governing photolytic cell injury processes.