Tracking of secretory vesicles of PC12 cells by total internal reflection fluorescence microscopy
Tracking of secretory vesicles of PC12 cells by total internal reflection fluorescence microscopy
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DOI：
10.1046/j.1365-2818.2003.01129.x
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发表时间：
2003-03
影响因子：
2
通讯作者：
Dm Yang;C.Chris Huang;H.‐Y. Lin;D. Tsai;L. Kao;Cheng Chi;C-C Lin-C
中科院分区：
文献类型：
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作者：
Dm Yang;C.Chris Huang;H.‐Y. Lin;D. Tsai;L. Kao;Cheng Chi;C-C Lin-C
全内反射荧光显微镜被用来检测质膜附近的细胞事件。研究了神经内分泌细胞系PC12活体细胞表面附近分泌小泡的行为。分泌囊泡通过增强的绿色荧光蛋白标记的Rab3A的过表达来标记，Rab3A是参与PC12细胞分泌囊泡与质膜融合的小G蛋白之一。对快速冷却电荷耦合器件相机使用常规荧光显微镜和全内反射荧光显微镜获得的图像进行了比较和分析。在小的消逝范围内(~lt；200 nm)，观察了PC12细胞在高K+刺激前后分泌囊泡的运动。检测一个囊泡相对于已经停靠在膜上的另一个囊泡的运动。全内反射荧光显微镜提供了一种新的光学方法来跟踪和分析细胞膜附近的胞吐事件和囊泡，而不会受到细胞其他部分信号的干扰。
Total internal reflection fluorescence microscopy is used to detect cellular events near the plasma membrane. Behaviours of secretory vesicles near the cell surface of living PC12 cells, a neuroendocrine cell line, are studied. The secretory vesicles are labelled by over‐expression of enhanced green fluorescent protein‐tagged Rab3A, one of the small G proteins involved in the fusion of secretory vesicles to plasma membrane in PC12 cells. Images acquired by a fast cooled charge‐coupled device camera using conventional fluorescence microscopy and total internal reflection fluorescence microscopy are compared and analysed. Within the small evanescent range (< 200 nm), the movements of the secretory vesicles of PC12 cells before and after stimulation by high K+ are examined. The movements of one vesicle relative to another already docked on the membrane are detected. Total internal reflection fluorescence microscopy provides a novel optical method to trace and analyse the exocytotic events and vesicle specifically near a cell membrane without interference of signals from other parts of the cell.