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Micromachines 2017, 8(2), 36;

Microfluidic Cell Cycle Analysis of Spread Cells by DAPI Staining

College of Environmental and Chemical Engineering, Institute of Microanalysis, Dalian University, Dalian 116622, China
School of Life Science and Biotechnology, Dalian University, Dalian 116622, China
Medical College, Institute of Microanalysis, Dalian University, Dalian 116622, China
Affiliated Zhongshan Hospital of Dalian University, Dalian 116622, China
Department of Chemistry and Pharmacy, Zhuhai College of Jilin University, Zhuhai 519041, China
This paper is an extended version of our paper published in the 2016 International Conference of Microfluidics, Nanofluidics and Lab-on-a-Chip (2016 ICMFLOC) was held in Dalian, China, 10–12 June 2016.
Authors to whom correspondence should be addressed.
Academic Editors: Yongxin Song, Junsheng Wang and Dongqing Li
Received: 5 December 2016 / Revised: 17 January 2017 / Accepted: 18 January 2017 / Published: 24 January 2017
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Single-cell cell cycle analysis is an emerging technique that requires detailed exploration of the image analysis process. In this study, we established a microfluidic single-cell cell cycle analysis method that can analyze cells in small numbers and in situ on a microfluidic chip. In addition, factors that influenced the analysis were carefully investigated. U87 or HeLa cells were seeded and attached to microfluidic channels before measurement. Cell nucleic DNA was imaged by 4′-6-diamidino-2-phenylindole (DAPI) staining under a fluorescent microscope and subsequently fluorescent intensities of the cell nuclei DNA were converted to depict histograms for cell cycle phases. DAPI concentration, microscopic magnification, exposure time and cell number were examined for optimal cell cycle analysis conditions. The results showed that as few as a few hundred cells could be measured by DAPI staining in the range of 0.4–0.6 μg/mL to depict histograms with typical cell cycle phase distribution. Microscopic magnification during image acquisition, however, could distort the phase distribution. Exposure time did not significantly affect the cell cycle analysis. Furthermore, cell cycle inhibitor rapamycin treatment changed the cell cycle phase distribution as expected. In conclusion, a method for microfluidic single-cell cell cycle analysis of spread cells in situ was developed. Factors such as dye concentration and microscopic magnification had more influence on cell cycle phase distribution. Further studies will focus on detail differentiation of cell cycle phases and the application of such a method for biological meanings. View Full-Text
Keywords: microfluidics; cell cycle; spread cell; DNA content microfluidics; cell cycle; spread cell; DNA content

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Sun, J.; Zhang, J.; Yang, H.; Wang, G.; Li, Y.; Zhang, X.; Chen, Q.; Lang, M.-F. Microfluidic Cell Cycle Analysis of Spread Cells by DAPI Staining. Micromachines 2017, 8, 36.

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