细胞大型内吞囊泡快速标记方法的改进和应用  

Improvement and application of a fast labeling system for bulk internalized vesicles

作　　者：戎叶 杜仲燕 肖桂凤[2] 娄绘芳[2] 吴航军[2] RONG Ye;DU Zhong-Yan;XIAO Gui-Feng;LOU Hui-Fang;WU Hang-Jun(Academy of Chinese Medical Sciences, Zhejiang Chinese Medical University, Hangzhou 311402, China;Zhejiang University School of Medicine, Hangzhou 310058, China)

机构地区：[1]浙江中医药大学中医药科学院,杭州311402 [2]浙江大学医学院,杭州310058

出　　处：《生理学报》2018年第3期287-293,共7页Acta Physiologica Sinica

基　　金：supported by the Natural Science Foundation of Zhejiang Province;China(No.LQ17C050001);the Fundamental Research Funds for the Central Universities of China(No.2015QNA7028);the Experimental Technology Research Program of Zhejiang University(No.SYB201510);the Research Foundation of Zhejiang Chinese Medical University;China(No.2018ZY36)

摘　　要：为了研究细胞内大型内吞囊泡的快速动态变化,本文借鉴了诱导神经元轴突诱导转向的方法,通过脉冲压力从微电极释放诱导因子,在细胞周围形成稳定的浓度梯度,诱导细胞产生胞饮泡,并通过局部染料释放进行快速标记和洗脱。同时利用活细胞成像技术,对胞饮泡标记过程以及标记后胞饮泡动态变化进行实时记录。该方法大大缩短了标记和洗脱的时间,从而实现了对细胞囊泡的快速标记和同步观察记录,可用于胞饮泡、溶酶体等细胞囊泡的动态标记和观察,是研究细胞大型内吞囊泡动态变化的一种实用且高效的手段。To study trafficking of bulk internalized vesicles such as macropinosome and lysosome in live cells, an efficient and convenient assay was established according to the axon turning assay. By injecting indicator or fluorescent dyes through a micropipette with air pressure into cell cultures to create a stable gradient around the micropipette tip, vesicles were indicated and labeled. With live cell imaging, the whole process was recorded. Without wash-out of fluorescent dyes and transferring, this assay is an effective, fast labeling system for bulk internalized vesicles, and can also be combined with imaging system.

关 键 词：囊泡标记 胞饮泡 溶酶体 活细胞成像 

分 类 号：Q31[生物学—遗传学] Q-331