Dynamic Quantitative Assays of Phagosomal Function

Q2 Immunology and Microbiology
Maria Podinovskaia, Brian C. VanderVen, Robin M. Yates, Sarah Glennie, Duncan Fullerton, Henry C. Mwandumba, David G. Russell
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引用次数: 26

Abstract

Much of the activity of the macrophage as an effector cell is performed within its phagocytic compartment. This ranges from the degradation of tissue debris as part of its homeostatic function to the generation of the superoxide burst as part of its microbicidal response to infection. We have developed a range of real‐time readouts of phagosomal function that enable these activities to be rigorously quantified. This unit contains descriptions of several of these assays assessed by different methods of quantitation, including a fluorescence resonance emission transfer (FRET) assay for phagosome/lysosome fusion measured by spectrofluorometry, a fluorogenic assay for the superoxide burst measured by flow cytometry, and a fluorogenic assay for bulk proteolysis measured by confocal microscopy. These assays illustrate both the range of parameters that can be quantified and the flexibility of instrumentation that can be exploited for their quantitation. Curr. Protoc. Immunol. 102:14.34.1‐14.34.14. © 2013 by John Wiley & Sons, Inc.

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吞噬体功能的动态定量测定
巨噬细胞作为效应细胞的大部分活动是在其吞噬室内进行的。其范围从组织碎片的降解作为其稳态功能的一部分到产生超氧化物爆发作为其对感染的杀微生物反应的一部分。我们开发了一系列吞噬体功能的实时读数,使这些活动能够严格量化。本单元包含了用不同的定量方法评估的几种检测方法的描述,包括用荧光光谱法测量吞噬体/溶酶体融合的荧光共振发射转移(FRET)检测,用流式细胞术测量超氧化物爆发的荧光检测,以及用共聚焦显微镜测量大量蛋白水解的荧光检测。这些分析既说明了可量化的参数范围,也说明了可用于其定量的仪器的灵活性。咕咕叫。Protoc。Immunol 102:14.34.1-14.34.14。©2013 by John Wiley &儿子,Inc。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Current Protocols in Immunology
Current Protocols in Immunology Immunology and Microbiology-Immunology
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