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

RONG Ye¹, DU Zhong-Yan¹, XIAO Gui-Feng², LOU Hui-Fang², WU Hang-Jun^2,*

¹Academy of Chinese Medical Sciences, Zhejiang Chinese Medical University, Hangzhou 311402, China；²Zhejiang University School of Medicine, Hangzhou 310058, China

Abstract

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.

Key words: vesicle labeling; macropinosome; lysosome; live cell imaging

Received: 2017-12-15　　Accepted: 2018-03-05

Corresponding author: 吴航军　　E-mail: wuhangjun@zju.edu.cn

DOI: 10.13294/j.aps.2018.0030

Citing This Article：

RONG Ye, DU Zhong-Yan, XIAO Gui-Feng, LOU Hui-Fang, WU Hang-Jun. Improvement and application of a fast labeling system for bulk internalized vesicles. Acta Physiol Sin 2018; 70 (3): 287-293 (in Chinese with English abstract).