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Clinical Chemistry 53: 2177-2185, 2007. First published October 25, 2007; 10.1373/clinchem.2007.092023
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(Clinical Chemistry. 2007;53:2177-2185.)
© 2007 American Association for Clinical Chemistry, Inc.

Automation and Analytical Techniques

Visual Recognition and Efficient Isolation of Apoptotic Cells with Fluorescent-Magnetic-Biotargeting Multifunctional Nanospheres

Er-Qun Song1, Guo-Ping Wang1, Hai-Yan Xie2, Zhi-Ling Zhang1, Jun Hu1, Jun Peng1, Dao-Cheng Wu3, Yun-Bo Shi4 and Dai-Wen Pang1,a

1 College of Chemistry and Molecular Sciences and State Key Laboratory of Virology, Wuhan University, Wuhan, Peoples Republic of China.
2 School of Life Science and Technology, Beijing Institute of Technology, Beijing, Peoples Republic of China.
3 School of Life Science and Technology, Xi’an Jiaotong University, Peoples Republic of China.
4 Section on Molecular Morphogenesis, Program on Cell Regulation and Metabolism, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD.

aAddress correspondence to this author at: College of Chemistry and Molecular Sciences and State Key Laboratory of Virology, Wuhan University, Wuhan 430072, Peoples Republic of China. Fax 86-27-6875-4067; e-mail dwpang{at}whu.edu.cn.

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.






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Copyright © 2007 by the American Association for Clinical Chemistry.