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引用本文:   张泽杰, 苏喜, 徐溢, 陈李. 多功能芯片对合成样本中肝癌细胞HepG2的测试研究. 分析化学, 2017, 45(11): 1589-1594. doi:  10.11895/j.issn.0253-3820.171092 [复制]

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

多功能芯片对合成样本中肝癌细胞HepG2的测试研究

通讯作者:  徐溢, xuyibbd@sina.com; 陈李, CL2009@cqu.edu.cn

收稿日期: 2017-07-13

接受日期: 2017-09-06

出版日期: 2017-11-20

基金项目: 本文系国家自然科学基金项目(No.21375156)、国家高技术研究发展计划(863)项目(No.2015AA021104)、重庆市前沿研究重点项目(No.cstc2015jcyjBX0010)和重庆市科学技术委员会社会民生科技创新项目(No.cstc2015shms zx00014)资助

Detection of HepG2 Cells in Artificial Samples by Multifunctional Microfluidic Chip

Corresponding author:  XU Yi, xuyibbd@sina.com; CHEN Li, CL2009@cqu.edu.cn

Received Date:  2017-07-13

Accepted Date:  2017-09-06

Published Date:  2017-11-20

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)

设计并制作了一种集多孔流分离(Multi-orifice flow fractionation,MOFF)技术与磁捕获技术于一体的用于特异性分离和捕获合成样本中肝癌细胞HepG2的多功能微流控细胞芯片。此芯片由玻璃基片和PDMS微通道盖片组成,PDMS盖片上含有3条进样通道、MOFF分离区和六边形腔体的细胞富集检测区。其中,MOFF分离区总长20 mm,由80组长度为0.18 mm、深度为50μm、收缩区域宽度为0.06 mm、扩张区域宽度为0.20 mm的半菱形收缩/扩张重复单元组成,每组收缩/扩张重复单元间的夹角为103.0°。实验以肝癌细胞HepG2-血细胞混悬液为样本;根据磁珠表面修饰c-Met抗体能与肝癌细胞HepG2特异性结合的原理,通过表面羧基化的磁珠、EDC(1 mg/mL)、NHS(1 mg/mL)和c-Met抗体制备了浓度为50μg/mL的免疫磁珠(Anti-MNCs)悬浮液。在样本流速为50μL/min条件下,利用外加磁场实现了血细胞合成样本中微量肝癌细胞HepG2的有效捕获;采用微波加热法以柠檬酸、硫脲为原料制备了用于荧光标记HepG2的碳量子点,在芯片上实现了血液中肝癌细胞HepG2的原位荧光可视化观测。对芯片检测区捕获到的HepG2进行了显微计数分析,对500μL血细胞(107 cell/mL)中含10个HepG2细胞的合成样本,捕获效率达到88.5%±6.7%(n=20)。结果表明,所设计的多模式多功能的微流控芯片具有良好的肿瘤细胞分离和检测功能。

关键词:   微流控芯片, 多孔流分离, 磁捕获, 荧光可视化, 肝癌细胞HepG2
Key words:   Microfluidic chip, Multi-orifice flow fractionation, Magnetic capture, Fluorescence visualization, HepG2
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多功能芯片对合成样本中肝癌细胞HepG2的测试研究

张泽杰, 苏喜, 徐溢, 陈李

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