多功能芯片对合成样本中肝癌细胞HepG2的测试研究

发布时间：2018-03-06 08:40

本文选题：微流控芯片　切入点：多孔流分离　出处：《分析化学》2017年11期 　论文类型：期刊论文

【摘要】：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Hep G2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Hep G2-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Hep G2特异性结合的原理,通过表面羧基化的磁珠、EDC(1 mg/m L)、NHS(1 mg/m L)和c-Met抗体制备了浓度为50μg/m L的免疫磁珠(AntiMNCs)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Hep G2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Hep G2的碳量子点,在芯片上实现了血液中肝癌细胞Hep G2的原位荧光可视化观测。对芯片检测区捕获到的Hep G2进行了显微计数分析,对500μL血细胞(107cell/m L)中含10个Hep G2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。
[Abstract]:A multifunctional microfluidic cell chip for specific isolation and capture of liver cancer cell line Hep G2 in synthetic samples was designed and fabricated, which integrates multi-orifice flow fractionation technique and magnetic capture technique. The microfluidic cell chip consists of glass substrate. And the PDMS microchannel capping plate consists of three sample injection channels, moff separation area and hexagonal cavity cell enrichment detection area. The total length of the PDMS is 20 mm, the length of 80 groups is 0.18 mm, the depth is 50 渭 m, and the width of contraction area is 0.06. A semirhombic contraction / dilatation repeating unit with a width of 0.20 mm in the expansion area, The angle of contraction / dilatation repeats in each group was 103.0 掳. The experiment took the suspension of Hep G2-blood cells as the sample, according to the principle that c-Met antibody modified by magnetic beads could specifically bind to Hep G2 of hepatoma cells. The suspension of 50 渭 g / mL immunomagnetic bead antiMNCswere prepared by carboxylated magnetic bead EDC1 mg/m mg/m L) and c-Met antibody. The flow rate of the sample was 50 渭 L / min. The effective capture of trace hepatoma cell Hep G2 in blood cell synthesis samples was realized by applying external magnetic field. Carbon quantum dots (QDs) for fluorescent labeling of Hep G2 were prepared by microwave heating method using citric acid and thiourea as raw materials. In situ fluorescence visualization of hepatoma cell Hep G2 in blood was carried out on the chip. The Hep G2 captured in the detection region of the microarray was counted and analyzed. The synthetic samples of 10 Hep G2 cells were obtained from the 500 渭 L blood cell line 107 cell / mL. The capture efficiency was 88.5% 卤6.7 and 20%. The results showed that the multi-mode and multi-function microfluidic chip had a good function of tumor cell separation and detection.
【作者单位】：重庆大学化学化工学院;重庆大学微纳系统及新材料技术国际研发中心;重庆大学新型微纳器件与系统技术重点学科实验室;重庆大学光电工程学院微系统研究中心;
【基金】：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.cstc2015jcyj BX0010) 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~~
【分类号】：O657.3;R735.7

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