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A prototypic microfluidic platform generating stepwise concentration gradients for real-time study of cell apoptosis
Authors:Wen Dai  Yizhe Zheng  Kathy Qian Luo  and Hongkai Wu
Institution:1Department of Chemistry, Hong Kong University of Science and Technology, Clean Water Bay, Kowloon, Hong Kong, China;2The NSNT Program, Hong Kong University of Science and Technology, Clean Water Bay, Kowloon, Hong Kong, China;3Division of Bioengineering, School of Chemical and Biomedical Engineering, Nanyang Technological University, 637457 Singapore, Singapore
Abstract:This work describes the development of a prototypic microfluidic platform for the generation of stepwise concentration gradients of drugs. A sensitive apoptotic analysis method is integrated into this microfluidic system for studying apoptosis of HeLa cells under the influence of anticancer drug, etoposide, with various concentrations in parallel; it measures the yellow fluorescent protein∕cyan fluorescent protein fluorescence resonance energy transfer (FRET) signal that responds to the activation of caspase-3, an indicator of cell apoptosis. Sets of microfluidic valves on the chip generate stepwise concentration gradient of etoposide in various cell-culture microchambers. The FRET signals from multiple chambers are simultaneously monitored under a fluorescent microscope for long-time observation and the on-chip results are compared with those from 96-well plate study and the methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay. The microfluidic platform shows several advantages including high-throughput capacity, low drug consumption, and high sensitivity.
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