Developing a microfluidic-based system to quantify cell capture efficiency  

作　　者：YANG Fan, GAO YuXin, ZHANG Yan, CHEN Juan & LONG Mian National Microgravity Laboratory and Center for Biomechanics and Bioengineering, Institute of Mechanics, Chinese Academy of Sciences, Beijing 100080, China 

出　　处：《Science China(Life Sciences)》2009年第2期173-181,共9页中国科学（生命科学英文版）

基　　金：Supported by the National Key Basic Research Program of China (Grant No. 2006CB910303);National Natural Science Foundation of China (Grant Nos. 30730032 and 10332060);National High-Tech Research and Development Program of China (Grant No. 2007AA02Z306);Chinese Academy of Sciences Grant (Grant No.2005-1-16)

摘　　要：Micro-fabrication technology has substantial potential for identifying molecular markers expressed on the surfaces of tissue cells and viruses. It has been found in several conceptual prototypes that cells with such markers are able to be captured by their antibodies immobilized on microchannel substrates and unbound cells are flushed out by a driven flow. The feasibility and reliability of such a microflu- idic-based assay, however, remains to be further tested. In the current work, we developed a microflu- idic-based system consisting of a microfluidic chip, an image grabbing unit, data acquisition and analysis software, as well as a supporting base. Specific binding of CD59-expressed or BSA-coupled human red blood cells (RBCs) to anti-CD59 or anti-BSA antibody-immobilized chip surfaces was quan- tified by capture efficiency and by the fraction of bound cells. Impacts of respective flow rate, cell concentration, antibody concentration and site density were tested systematically. The measured data indicated that the assay was robust. The robustness was further confirmed by capture efficiencies measured from an independent ELISA-based cell binding assay. These results demonstrated that the system developed provided a new platform to effectively quantify cellular surface markers effectively, which promoted the potential applications in both biological studies and clinical diagnoses.Micro-fabrication technology has substantial potential for identifying molecular markers expressed on the surfaces of tissue cells and viruses. It has been found in several conceptual prototypes that cells with such markers are able to be captured by their antibodies immobilized on microchannel substrates and unbound cells are flushed out by a driven flow. The feasibility and reliability of such a microflu- idic-based assay, however, remains to be further tested. In the current work, we developed a microflu- idic-based system consisting of a microfluidic chip, an image grabbing unit, data acquisition and analysis software, as well as a supporting base. Specific binding of CD59-expressed or BSA-coupled human red blood cells (RBCs) to anti-CD59 or anti-BSA antibody-immobilized chip surfaces was quan- tified by capture efficiency and by the fraction of bound cells. Impacts of respective flow rate, cell concentration, antibody concentration and site density were tested systematically. The measured data indicated that the assay was robust. The robustness was further confirmed by capture efficiencies measured from an independent ELISA-based cell binding assay. These results demonstrated that the system developed provided a new platform to effectively quantify cellular surface markers effectively, which promoted the potential applications in both biological studies and clinical diagnoses.

关 键 词：CELL SURFACE MARKER microfluidic CAPTURE EFFICIENCY 

分 类 号：Q2-3[生物学—细胞生物学]