采用多色流式细胞术分析评估活细胞中的线粒体含量和功能。

Evaluation of Mitochondria Content and Function in Live Cells by Multicolor Flow Cytometric Analysis.

机构信息

Institute of Microbiology and Immunology, National Yang-Ming University, Taipei, Taiwan.

出版信息

Methods Mol Biol. 2021;2276:203-213. doi: 10.1007/978-1-0716-1266-8_15.

Abstract

To evaluate how a cell responds to the external stimuli, treatment, or alteration of the microenvironment, the quantity and quality of mitochondria are commonly used as readouts. However, it is challenging to apply mitochondrial analysis to the samples that are composed of mixed cell populations originating from tissues or when multiple cell populations are of interest, using methods such as Western blot, electron microscopy, or extracellular flux analysis.Flow cytometry is a technique allowing the detection of individual cell status and its identity simultaneously when used in combination with surface markers. Here we describe how to combine mitochondria-specific dyes or the dyes targeting the superoxide produced by mitochondria with surface marker staining to measure the mitochondrial content and activity in live cells by flow cytometry. This method can be applied to all types of cells in suspension and is particularly useful for analysis of samples composed of heterogeneous cell populations.

摘要

为了评估细胞对外界刺激、处理或微环境改变的反应，线粒体的数量和质量通常被用作检测指标。然而，当涉及到由组织来源的混合细胞群体或当多个细胞群体为研究对象时，使用Western blot、电子显微镜或细胞外通量分析等方法来进行线粒体分析是具有挑战性的。流式细胞术是一种技术，当与表面标志物结合使用时，可以同时检测单个细胞的状态及其身份。在这里，我们描述了如何将线粒体特异性染料或靶向线粒体产生的超氧化物的染料与表面标志物染色结合使用，通过流式细胞术测量活细胞中的线粒体含量和活性。该方法可应用于所有悬浮细胞类型，对于分析由异质细胞群体组成的样本尤其有用。