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通过流式细胞术和图像细胞术对细胞凋亡进行多参数分析。

Multiparametric analysis of apoptosis by flow and image cytometry.

作者信息

Telford William G, Komoriya Akira, Packard Beverly Z

机构信息

Experimental Transplantation and Immunology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.

出版信息

Methods Mol Biol. 2004;263:141-60. doi: 10.1385/1-59259-773-4:141.

Abstract

Flow cytometric assays for apoptosis are now in widespread use. The multiparametric nature of flow cytometry allows multiple assays for several apoptotic characteristics to be combined in a single sample, providing a powerful tool for elucidating the complex progression of apoptotic death in a variety of cell types. This chapter describes one such assay, allowing simultaneous analysis of caspase activation, annexin V binding to "flipped" phosphatidylserine residues and membrane permeability to DNA binding dyes. This multidimensional approach to analyzing apoptosis provides far more information than single-parameter assays that provide only an ambiguous "percent apoptotic" result, given that multiple early, intermediate, and late apoptotic stages can be visualized simultaneously. This multiparametric approach is also amenable to a variety of flow cytometric instrumentation, both old and new.

摘要

用于凋亡检测的流式细胞术目前已被广泛应用。流式细胞术的多参数特性使得在单个样本中能够结合对多种凋亡特征进行多种检测,为阐明各种细胞类型中凋亡死亡的复杂进程提供了一个强大的工具。本章介绍了这样一种检测方法,它能够同时分析半胱天冬酶激活、膜联蛋白V与“翻转”的磷脂酰丝氨酸残基结合以及细胞膜对DNA结合染料的通透性。这种用于分析凋亡的多维方法所提供的信息远比单参数检测多得多,单参数检测只能给出一个模糊的“凋亡百分比”结果,因为多个早期、中期和晚期凋亡阶段能够同时可视化。这种多参数方法也适用于各种新旧流式细胞仪。

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