Citation: ZHANG Ze-Jie, SU Xi, XU Yi, CHEN Li. Detection of HepG2 Cells in Artificial Samples by Multifunctional Microfluidic Chip[J]. Chinese Journal of Analytical Chemistry, ;2017, 45(11): 1589-1594. doi: 10.11895/j.issn.0253-3820.171092 shu

Detection of HepG2 Cells in Artificial Samples by Multifunctional Microfluidic Chip

ZHANG Ze-Jie ^2,4, ,
SU Xi ^1,2,4 ,
XU Yi ^1,2,3,4 ,
CHEN Li ^1,3,4

1.
Key Disciplines Lab of Novel Micro-nano Devices and System Technology, Chongqing University, Chongqing 400044, China;
2.
School of Chemistry and Chemical Engineering, Chongqing University, Chongqing 400044, China;
3.
School of Optoelectronic Engineering, Research Center of Microsystem, Chongqing University, Chongqing 400044, China;
4.
International R & D center of Micro-nano Systems and New Materials Technology, Chongqing University, Chongqing 400044, China

Received Date: 13 July 2017
Revised Date: 6 September 2017

Fund Project: This work was supported by the National Natural Science Foundation of China (No. 21375156), the National High Technology Research and Development Program of China (863 program) (No. 2015AA021104), the Frontier Research Key Projects of Chongqing Science and Technology Committee, China (No. cstc2015jcyjBX0010), and the Scientific and Technical Innovation Projects for People's Livelihood of Chongqing Science and Technology Committee, China (No. cstc2015shmszx00014).

A multi-functional microfluidic chip with multi-orifice flow fractionation (MOFF) and magnetic capture technique was developed to specifically separate and capture the HepG2 cells in artificial samples. The chip contained a glass substrate and a polydimethylsiloxane (PDMS) microchannel cover plate. The PDMS cover plate consisted of 3 injection channels of 10-mm-long, a MOFF separation zone and a hexagonal cavity cell enrichment-detection zone. Among which, the MOFF separation zone had a total length of 20 mm and was consisted of 80 semi-rhombic shrinkage/expansion units with a length of 0.18 mm, a depth of 50 μm, a shrinkage area width of 0.06 mm, and an expansion area of 0.20 mm. The angle between each group of shrinkage/expansion units was 103.0°. In this experiment, HepG2-blood cell suspension was used as the sample. Based on the principle that the magnetic bead surface-modified c-Met antibody could specifically bind to HepG2 cells, an immunomagnetic bead (Anti-MNCs) suspension at a concentration of 50 μg/mL was prepared by surface carboxylated beads, EDC (1 mg/mL), NHS (1 mg/mL) and c-Met antibody. Under the optimized flow rate (50 μL/min), a few HepG2 in suspension samples were efficiently captured at the detection zone of chip via a magnetic field; the carbon quantum dots were prepared by microwave heating with citric acid and thiourea to label HepG2 cells which achieved in-situ fluorescence visualization of captured HepG2. Cells captured in the chip detection area were counted by microscope. The capture rate of HepG2 cells was 88.5%±6.7% (10⁶ blood cells and 10 HepG2 cells per 500 μL). The results demonstrated that the developed multifunctional microfluidic chip may serve as a promising tool for separation and capture of tumour cells.
- Microfluidic chip,
- Multi-orifice flow fractionation,
- Magnetic capture,
- Fluorescence visualization,
- HepG2
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