高通量免疫反应微流控细胞芯片.PDFVIP

Vol. 4 No.6/ Jun. 010 高通量免疫反应微流控细胞芯片 High-Throughput Multi-antigen Immunoassays on a Microfluidic chip 张宝月 宋惠雪 陈坦 孙宝国 王战会 摘 要 传统基于细胞的实验如免疫反应主要采用96-孔板、培养瓶、载玻片等方法。这些方法往往操作烦琐、消耗试剂量 大、需要细胞数多，本研究研制一种微流控阵列细胞培养芯片，集成细胞培养、冲洗、固定、抗原-抗体反应等实验步骤。 显示了其自动化操作及高通量免疫反应的能力。借助此微流控芯片平台，每个免疫反应仅需几十到几百个细胞及纳升级的 反应试剂量。有效节省细胞及试剂，尤其是数量稀少的干细胞。此微流控细胞芯片平台通过同时检测人脐带血间充质干细 胞表面分子：CD34, CD44, CD45, CD73,CD105, HLA-DR的表达情况以证明。微流控芯片荧光免疫反应整个实验操作都 在微流控细胞芯片系统上完成，通过可控的流体实现对细胞悬液、固定液、封闭液、抗体溶液的进样及细胞的清洗，最后 经荧光显微镜成像采集荧光信号。证明了微流控芯片细胞培养系统的可行性，为建立一个基于细胞培养的微型化、集成 化、自动化、便携化微流控生物实验技术平台打下基础。 关键词 微流控技术； 细胞培养；免疫荧光反应；软光刻技术 ABSTRACT　Traditional cell-based assays such as cell immunoassay that utilizes plastic (chamber slides, dishes, microtiter plates), Magnetic bead, enzyme-linked immunsorbent assays (ELISA) [1], FACS cell sorting is labor intensive, time consuming, and requires a large numbers of cells or reagents. In this report, a microfluidic device integrated with cell culture, washing, fixation, and antigen-antibody reaction is presented for high-throughput immunoassay. Using this microfluidic device, each assay can be performed on a small number of cells and nanolitre or picolitre of reagents, this is particularly beneficial for rare or expensive cell types such as stem cells, or flow sorted cell populations. The capability of the microfluidic device was demonstrated for seeding human umbilical cord blood mesenchymal stem cells (UC- MSCs) in chambers and detecting the expression of surface markers (CD34, CD44, CD45, CD73, CD105, HLA-DR) by immunofluorescence assay. KEYWORDS Microfluidic Technology; Cell Culture; Immunefluorescence; Soft Lithography 1 Introduction AMMALIAN cell culture has played a major role research, there is still no single marker that uniquely Min the development of