Quantifying autophagy: Measuring LC3 puncta and autolysosome formation in cells using multispectral imaging flow cytometry
Quantifying autophagy: Measuring LC3 puncta and autolysosome formation in cells using multispectral imaging flow cytometry
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DOI：
10.1016/j.ymeth.2016.05.022
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发表时间：
2017-01-01
期刊：
影响因子：
4.8
通讯作者：
Pugsley, Haley R.
中科院分区：
文献类型：
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作者：
Pugsley, Haley R.
多光谱成像流式细胞术的使用由于其定量能力、高通量能力、多路复用潜力及其获取每个细胞的图像的能力而越来越受欢迎。自噬是在生长和分化过程中积累的功能失调的细胞器和细胞组分通过溶酶体降解并再循环的过程。在自噬过程中，细胞质LC 3被加工并募集到自噬体膜;自噬体然后与溶酶体融合以形成自溶体。因此，经历自噬的细胞可以通过可视化荧光标记的LC 3斑点和/或荧光标记的LC 3和溶酶体标志物的共定位来鉴定。多光谱成像流式细胞术能够收集大量细胞的图像，并以客观、定量和统计稳健的方式评估自噬。这篇综述将研究四种主要的方法，已被用来测量自噬通过多光谱成像流式细胞术。(C)2016作者爱思唯尔公司出版
The use of multispectral imaging flow cytometry has been gaining popularity due to its quantitative power, high throughput capabilities, multiplexing potential and its ability to acquire images of every cell. Autophagy is a process in which dysfunctional organelles and cellular components that accumulate during growth and differentiation are degraded via the lysosome and recycled. During autophagy, cytoplasmic LC3 is processed and recruited to the autophagosomal membranes; the autophagosome then fuses with the lysosome to form the autolysosome. Therefore, cells undergoing autophagy can be identified by visualizing fluorescently labeled LC3 puncta and/or the co-localization of fluorescently labeled LC3 and lysosomal markers. Multispectral imaging flow cytometry is able to collect imagery of large numbers of cells and assess autophagy in an objective, quantitative, and statistically robust manner. This review will examine the four predominant methods that have been used to measure autophagy via multispectral imaging flow cytometry. (C) 2016 The Author. Published by Elsevier Inc.