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[微珠、纳米珠与细胞计数:在细胞及生物分子分析与纯化中的应用]

[Microbeads, nanobeads and cytometry: applications to the analysis and purification of cells and biomolecules].

作者信息

Lizard G, Duvillard L, Wedemeyer N, Muller C, Ghiringhelli F, Cesbron A, Poncelet P, Gallet F, Kahn E, Gambert P, Göhde W

机构信息

Laboratoire de biochimie médicale, CHU/hôpital du Bocage, Inserm U498, IFR 100, BP 77908, 21079 cedex, Dijon, France.

出版信息

Pathol Biol (Paris). 2003 Sep;51(7):418-27. doi: 10.1016/s0369-8114(03)00127-5.

Abstract

Nano and microspheres are important tools in cytometry. They have been used in first to optimize fluorescent signals detected by flow cytometry and to evaluate phagocytosis. Some antigens were also detected by using nanospheres covalently coupled to antibodies. Specifically dedicated microspheres are now widely used for antigenic quantitation by flow cytometry, and magnetic nano and micropheres are very usefull for cellular and molecular purifications. To date, analytical methods based on the use of microspheres are developed to detect proteins, nucleic acids, and ions. To this end, antibodies, oligonucleotides, or chelating agents are bound to microspheres characterized by different fluorescences. The applications of these multiplexed microspheres assays allow to identify and quantify simultaneously some macromolecules and ions, but they also permit to analyze enzymatic activities and to perform polymorphism analyses. With microspheres used as reactive support, molecular analyses are therefore possible by flow cytometry. Nano and microspheres are also usefull tools for calibration in confocal microscopy as well as for micromanipulations of biomolecules and of living cells. Inovative methods based on the use of nano and microspheres are expected in the fields of biology, medicine, food industry, and environmental sciences.

摘要

纳米球和微球是细胞计数中的重要工具。它们首先被用于优化流式细胞术检测的荧光信号以及评估吞噬作用。一些抗原也通过使用与抗体共价偶联的纳米球来检测。专门设计的微球现在广泛用于流式细胞术的抗原定量,磁性纳米球和微球对于细胞和分子纯化非常有用。迄今为止,基于微球使用的分析方法已被开发用于检测蛋白质、核酸和离子。为此,抗体、寡核苷酸或螯合剂与具有不同荧光特性的微球结合。这些多重微球分析的应用不仅可以同时识别和定量一些大分子和离子,还可以分析酶活性并进行多态性分析。因此,通过将微球用作反应支持物,利用流式细胞术进行分子分析成为可能。纳米球和微球也是共聚焦显微镜校准以及生物分子和活细胞微操作的有用工具。预计在生物学、医学、食品工业和环境科学领域会出现基于纳米球和微球使用的创新方法。

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