title: Laminar flow mediated continuous single-cell analysis on a novel poly(dimethylsiloxane) microfluidic chip.
abstract: A novel microfluidic chip with simple design, easy fabrication and low cost, coupled with high-sensitive laser induced fluorescence detection, was developed to provide continuous single-cell analysis based on dynamic cell manipulation in flowing streams. Making use of laminar flows, which formed in microchannels, single cells were aligned and continuously introduced into the sample channel and then detection channel in the chip. In order to rapidly lyse the moving cells and completely transport cellular contents into the detection channel, the angle of the side-flow channels, the asymmetric design of the channels, and the number, shape and layout of micro-obstacles were optimized for effectively redistributing and mixing the laminar flows of single cells suspension, cell lysing reagent and detection buffer. The optimized microfluidic chip was an asymmetric structure of three microchannels, with three microcylinders at the proper positions in the intersections of channels. The microchip was evaluated by detection of anticancer drug doxorubicin (DOX) uptake and membrane surface P-glycoprotein (P-gp) expression in single leukemia K562 cells. An average throughput of 6-8 cells min(-1) was achieved. The detection results showed the cellular heterogeneity in DOX uptake and surface P-gp expression within K562 cells. Our researches demonstrated the feasibility and simplicity of the newly developed microfluidic chip for chemical single-cell analysis.
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Ministry of Education), and School of Pharmaceutical Sciences, Wuhan University, Wuhan 430071, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Deng
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Ministry of Education), and School of Pharmaceutical Sciences, Wuhan University, Wuhan 430071, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Tian
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Wuhan University, Wuhan 430072, China.
authors.forename: Xu
authors.initials: X
authors.lastname: Yu
authors.affiliation: Department of Anatomy and Embryology, School of Medicine, Wuhan University, Wuhan 430071, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Song
authors.affiliation: Department of Engineering Science and Mechanics, The Pennsylvania State University, University Park, PA 16802, USA.
authors.forename: Feng
authors.initials: F
authors.lastname: Guo
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Ministry of Education), and School of Pharmaceutical Sciences, Wuhan University, Wuhan 430071, China. Electronic address: yuxiuxiao2011@whu.edu.cn.
authors.forename: Yuxiu
authors.initials: Y
authors.lastname: Xiao
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Wuhan University, Wuhan 430072, China. Electronic address: zlzhang@whu.edu.cn.
authors.forename: Zhiling
authors.initials: Z
authors.lastname: Zhang
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism; D001692:Biological Transport; D004129:Dimethylpolysiloxanes / Q000737:chemistry*; D004317:Doxorubicin / Q000378:metabolism; D004867:Equipment Design; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D020014:K562 Cells; D046210:Microfluidic Analytical Techniques / Q000191:economics / Q000295:instrumentation / Q000379:methods*; D059010:Single-Cell Analysis / Q000191:economics / Q000295:instrumentation / Q000379:methods*; D013997:Time Factors
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D004129:Dimethylpolysiloxanes; C013830:baysilon; D004317:Doxorubicin
keywords: Doxorubicin; Laminar flow; Microfluidic chip; P-glycoprotein; Single-cell analysis