化学学报 ›› 2014, Vol. 72 ›› Issue (1): 69-74.DOI: 10.6023/A13080902 上一篇    下一篇

研究论文

简易型微流控芯片捕获循环肿瘤细胞的研究

高菊逸a,b, 杜晶辉a,b, 张望a,b, 张宝月c, 刘宗彬c, 陈艳c, 徐小平b   

  1. a 南昌大学研究生院医学部 南昌 330006;
    b 香港大学深圳医院 深圳 518053;
    c 中国科学院深圳先进技术研究院 深圳 518055
  • 收稿日期:2013-08-31 出版日期:2014-01-14 发布日期:2013-10-30
  • 通讯作者: 徐小平,E-mail:jyoxu@163.com;Tel.:13828787979 E-mail:jyoxu@163.com
  • 基金资助:

    项目受深圳发展改革委员会项目[(2012)385];国家自然科学基金(No. 61106128)和深圳市科技计划项目(No. 201101004)资助.

A Simple Fabricated Microfluidic Chip for Rapid Capture of Circulating Tumor Cells

Gao Juyia,b, Du Jinghuia,b, Zhang Wanga,b, Zhang Baoyuec, Liu Zongbinc, Chen Yanc, Xu Xiaopingb   

  1. a Medical Department of Nanchang University Graduate School, Nanchang 330006;
    b Hong Kong University Shenzhen Hospital, Shenzhen 518053;
    c Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055
  • Received:2013-08-31 Online:2014-01-14 Published:2013-10-30
  • Supported by:

    Project supported by the Shenzhen Development and Reform Commission Project (Grant 2012-385), the National Natural Science Foundation of China (No. 61106128), Shenzhen Science and Technology Plan Project (No. 201101004).

本研究以循环肿瘤细胞(乳腺肿瘤细胞)为研究对象,以聚二甲基硅氧烷(PDMS)、双面粘性薄膜(DSA)、玻片为原材料,采用激光雕刻技术制作微流控芯片,结合巯基-马来酰亚胺基团硅烷化偶联法和免疫荧光技术进行芯片内捕获检测实验,并使用外周血肿瘤细胞来验证此微流控芯片的实用性,使用具有高速摄像功能的荧光显微镜进行镜下观察及拍摄. 成功构建了一种简易型微流控芯片系统,利用此系统可实现对乳腺肿瘤细胞(92±3)%的捕获率,对外周血肿瘤细胞(88±3)%的捕获率,而且芯片的制作工艺简单,对实验仪器要求低,1 min内即可制作完成,简化了制作过程,弥补了传统光刻工艺复杂繁琐的不足,为临床检测疾病的发生与发展提供了新的研究方向.

关键词: 微流控芯片, 捕获, 循环肿瘤细胞, 简易, 双面粘性薄膜

Due to their portability, affordability and high sensitivity, microfluidic systems have become promising technologies to develop point-of-care (POC) diagnostics. A major challenge in microfluidic system fabrication is to integrate multiple microfluidic components in a simple way and construct microfluidic systems in a rapid, inexpensive manner. Here we investigated the design and fabrication of rapid capture of circulating tumor cells (CTCs) without the need for a cleanroom facility. The device contains is composed of a polydimethylsiloxane (PDMS) layer and a double-sided adhesive film (DSA). The microfluidic channels on the DSA film were simply fabricated using a laser prototyping technique. PDMS was chosen as the support layer to facilitate sample injection and avoid leakage. The microfluidic chip fabrication takes less than one minute without employing expensive lithographic methods. Design changes of fluidic channel can be made by ProE software, and which is applicable to rapid prototyping of microfluidic devices. The PDMS base and a glass cover slip were then assembled via the DSA. Before assembling the chip, glass cover was cleaned with ethanol using sonication. Then, it was washed with distilled water and dried under nitrogen gas. After cleaning steps, the glass cover was plasma treated for 60 seconds (100 mW, 1% O2, 60 s). Next, specific antibody was pre-immobilized on the bottom of the microchannel and then bound with FITC-labeled detection antibodies to generate fluorescent signals. Then, the inlet with cell suspension solution was connected to a syringe pump which controlled the fluid velocity, flow rates was adjusted from 2 to 8 μL/min. Using this device, a high tumor cell capture efficiency of 92% was achieved at a flow velocity of 2 μL/min. In this paper, a new technique is described to reduce fabrication time and cost, and shows considerable promise for the translation to a rapid point-of-care diagnostic device for the detection of CTCs.

Key words: microfluidic chip, capture, circulating tumor cells, simple, double-sided adhesive film