Visual recognition and efficient isolation of apoptotic cells with a fluorescent-magnetic-biotargeting multifunctional nanospheres
Visual recognition and efficient isolation of apoptotic cells with a fluorescent-magnetic-biotargeting multifunctional nanospheres
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利用荧光磁生物靶向多功能纳米球视觉识别和有效分离凋亡细胞
DOI：
10.1373/clinchem.2007.092023
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发表时间：
2007-12-01
影响因子：
9.3
通讯作者：
Pang, Dai-Wen
中科院分区：
文献类型：
--
作者：
Song, Er-Qun;Wang, Guo-Ping;Pang, Dai-Wen
背景：荧光-磁-生物靶向多功能纳米球可能在生物分析、生物医学和临床诊断中找到重要的应用。我们一直在开发用于生物医学的多功能纳米球。方法：我们将亲和素共价偶联到荧光磁双功能纳米球表面，构建荧光磁生物靶向三功能纳米球，并分析这些三功能纳米球识别和分离凋亡细胞的功能和特异性，这些三功能纳米球标记有生物素化膜联蛋白V，可以识别暴露在凋亡细胞表面的磷脂酰丝氨酸。结果：多功能纳米微球结合碘化丙啶染色核DNA可用于鉴定细胞凋亡过程的不同阶段。此外，我们证明，暴露于紫外光诱导的凋亡细胞可以简单地用磁铁从活细胞中分离出来，效率至少为80%；这些细胞可以很容易地用荧光显微镜观察到。结论：荧光-磁-生物靶向三功能纳米球可作为快速识别、磁富集和分选、同时识别不同类型细胞的有力工具。(C) 2007年美国临床化学学会。
Background: Fluorescent-magnetic-biotargeting multifunctional nanospheres are likely to find important applications in bioanalysis, biomedicine, and clinical diagnosis. We have been developing such multifunctional nanospheres for biomedical applications.Methods: We covalently coupled avidin onto the surfaces of fluorescent-magnetic bifunctional nanospheres to construct fluorescent-magnetic-biotargeting trifunctional nanospheres and analyzed the functionality and specificity of these trifunctional nanospheres for their ability to recognize and isolate apoptotic cells labeled with biotinylated annexin V, which recognizes phosphatidylserine exposed on the surfaces of apoptotic cells.Results: The multifunctional nanospheres can be used in combination with propidium iodide staining of nuclear DNA to identify cells at different phases of the apoptotic process. Furthermore, we demonstrate that apoptotic cells induced by exposure to ultraviolet light can be isolated simply with a magnet from living cells at an efficiency of at least 80%; these cells can then be easily visualized with a fluorescence microscope.Conclusions: Our results show that fluorescent-magnetic-biotargeting trifunctional nanospheres can be a powerful tool for rapidly recognizing, magnetically enriching and sorting, and simultaneously identifying different kinds of cells. (C) 2007 American Association for Clinical Chemistry.