激光生物学报, 2015, 24 (4): 326, 网络出版: 2015-11-30
微激光照射肥大细胞对胞内钙离子浓度的影响
The Effect of Micro-laser on Intracellular Free Calcium Concentration of Mast Cell
生物光学 细胞内钙离子浓度 微激光 肥大细胞 激光扫描共聚焦显微镜 biological optics intracellular calcium concentration ([Ca2+] i ) micro-laser mast cell laser scanning confocal microscopy(LSCM)
摘要
目的:研究850 nm波长微激光对肥大细胞照射后胞内钙离子浓度的影响。方法:实验前12 h,将肥大细胞接种于激光共聚焦专用培养皿中,然后用D- Hank’s液洗涤3次,加入2 mL D-Hank’s液,按照照射时间不同共分六组(Control, 1 min, 2 min, 2.5 min, 3 min, 4 min),其中,空白对照组不进行激光照射处理;激光照射后,弃去D- Hank’s液,加入含有Fluo-3/AM的D- Hank’s液1 mL,放入培养箱中孵育30 min后,用D- Hank’s液洗涤3次去除胞外多余的Fluo-3/AM,再加入含有10%FBS的D- Hank’s液2 mL,置激光扫描共聚焦显微镜检测。结果:空白对照组中细胞内未见荧光,说明此时胞内无钙离子,但从1 min开始在细胞中发现荧光,而且发现随着照射时间的加长,其荧光强度越来越强,这可能与微激光照射时间越长从而刺激胞内出现更多的钙离子有关。从上面的实验说明850 nm微激光能作为仿灸仪器的光源。
Abstract
Objective:To investigate the effects of micro-laser irradiation on intracellular calcium concentration ([Ca2+]i) of mast cell. Methods:12 hours before the experiment, The mast cells were seeded into paltes, which were used on the laser scanning confocal microscopy(LSCM). Then the cells were washed three times with D-hank’s solution, and added 2 mL D-hank’s solution to each group. There were six groups, which were irradiated at different times (Control, 1 min, 2 min, 2.5 min, 3 min, 4 min) by micro-laser 850 nm.The control group was irradiated 0 min. After micro-laser irradiation, the cells were removed the D-hank’s solution, and loaded with 1 mL Fluo-3/AM at 37°C for 30 min. The cells were washed again for another three times with D-hank’s solution to remove the unloaded Fluo-3/AM in the medium, and then added 2 mL D-hank’s solution with 10% FBS. The images of Ca2+ concentration in each group cells were monitored by LCSM. Result:The results have shown that there were no fluorescence in the control group. It clearly indicated that there were no Ca2+ in the control group cells. But in the irradiated 1 min group, there was significant fluorescence. Following the time of irradiation increasing, the fluorescence intensity were enhanced. The intracellular calcium concentration become higher, which may be related to the micro-laser irradiation time increase. It has shown that the 850 nm micro-laser could be the optical source of moxibustion instrument.Key words:
钟会清, 杨辉, 周艳, 郭周义, 黄汉传, 叶丙刚, 倪艺榕, 熊红莲, 金梅, 吴秀丽, 苏成康, 龙佳, 林锦. 微激光照射肥大细胞对胞内钙离子浓度的影响[J]. 激光生物学报, 2015, 24(4): 326. ZHONG Huiqing, YANG Hui, ZHOU Yan, GUO Zhouyi, HUANG hanchuan, YE Bingang, NI Yirong, XIONG Honglian, JIN Mei, WU Xiuli, SU Chengkang, LONG Jia, LIN Jin. The Effect of Micro-laser on Intracellular Free Calcium Concentration of Mast Cell[J]. Acta Laser Biology Sinica, 2015, 24(4): 326.