期刊
AUTOPHAGY
卷 19, 期 1, 页码 338-351出版社
TAYLOR & FRANCIS INC
DOI: 10.1080/15548627.2022.2065617
关键词
Airyscan microscopy; autophagy; CellProfiler; lipid droplets; SQSTM1; p62; WIPI1
类别
This paper presents a method for single cell-based analysis of autophagy using open source CellProfiler software. The method utilizes fluorescence microscopy to detect autophagy-related proteins and provides a reliable and automated analysis of the number and size of recognized puncta. The method can be easily adapted for different autophagy markers or cell types and is suitable for high-throughput studies.
Single cell-based analysis of macroautophagy/autophagy is largely achieved through the use of fluorescence microscopy to detect autophagy-related proteins that associate with autophagic membranes and therefore can be quantified as fluorescent puncta. In this context, an automated analysis of the number and size of recognized puncta is preferable to a manual count, because more reliable results can be generated in a short time. Here we present a method for open source CellProfiler software-based analysis for quantitative autophagy assessments using GFP-tagged WIPI1 (WD repeat domain, phosphoinositide interacting 1) images acquired with Airyscan or confocal laser-scanning microscopy. The CellProfiler protocol is provided as a ready-to-use software pipeline, and the creation of this pipeline is detailed in both text and video formats. In addition, we provide CellProfiler pipelines for endogenous SQSTM1/p62 (sequestosome 1) or intracellular lipid droplet (LD) analysis, suitable to assess forms of selective autophagy. All protocols and software pipelines can be quickly and easily adapted for the use of alternative autophagy markers or cell types, and can also be used for high-throughput purposes.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据