胞吐
内吞循环
小泡
突触小泡
内吞作用
荧光显微镜
化学
荧光
体内吞
内体
细胞生物学
生物物理学
分泌泡
生物
生物化学
细胞内
分泌物
膜
细胞
光学
物理
作者
Michael A. Gaffield,William J. Betz
出处
期刊:Nature Protocols
[Springer Nature]
日期:2006-12-01
卷期号:1 (6): 2916-2921
被引量:229
标识
DOI:10.1038/nprot.2006.476
摘要
FM dyes have been used to label and then monitor synaptic vesicles, secretory granules and other endocytic structures in a variety of preparations. Here, we describe the general procedure for using FM dyes to study endosomal trafficking in general, and synaptic vesicle recycling in particular. The dye, dissolved in normal saline solution, is added to a chamber containing the preparation to be labeled. Stimulation evokes exocytosis, and compensatory endocytosis that follows traps FM dye inside the retrieved vesicles. The extracellular dye is then washed from the chamber, and labeled endocytic structures are examined with a fluorescence microscope. Fluorescence intensity provides a direct measure of the labeled vesicle number, a good measure of the amount of exocytosis. If the preparation is stimulated again, without dye in the chamber, dimming of the preparation provides a measure of exocytosis of labeled vesicles. With a synaptic preparation on hand, this protocol requires 1 day.
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