title: Pneumatic microfluidics-based multiplex single-cell array.
abstract: Large-scale single-cell arrays are urgently required for current high-throughput screening of cell function and heterogeneity. However, the rapid and convenient generation of large-scale single-cell array in a multiplex and universal manner is not yet well established. In this paper, we report a simple and reliable method for the generation of a single-cell array by combining pneumatic microvalve arrays (PμVAs) and hydrodynamic single-cell trapping sites in a single microfluidic device. The PμVAs, which can be precisely controlled by actuated pressures, were designed to guide multiple types of cells being trapped in the corresponding single-cell trapping sites located in the fluidic channel. According to the theoretical demonstration and computational simulation, we successfully realized a multiplex single-cell array with three different types of cells by a step-by-step protocol. Furthermore, the analysis of cellular esterase heterogeneity of the three types of cells was concurrently implemented in the device as a proof-of-concept experiment. All the results demonstrated that the method developed in the current study could be applied for the generation of large-scale single-cell array with multiple cell types, which would be also promising and helpful for single-cell-based high-throughput drug test, multipurpose immunosensor and clinical diagnosis.
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
authors.forename: Lei
authors.initials: L
authors.lastname: Zhao
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
authors.forename: Chao
authors.initials: C
authors.lastname: Ma
authors.affiliation: College of Science, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
authors.forename: Shaofei
authors.initials: S
authors.lastname: Shen
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
authors.forename: Chang
authors.initials: C
authors.lastname: Tian
authors.affiliation: College of Science, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
authors.forename: Juan
authors.initials: J
authors.lastname: Xu
authors.affiliation: College of Science, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
authors.forename: Qin
authors.initials: Q
authors.lastname: Tu
authors.affiliation: College of Science, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
authors.forename: Tianbao
authors.initials: T
authors.lastname: Li
authors.affiliation: College of Science, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
authors.forename: Yaolei
authors.initials: Y
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, PR China; College of Science, Northwest A&F University, Yangling, Shaanxi 712100, PR China. Electronic address: jywang@nwsuaf.edu.cn.
authors.forename: Jinyi
authors.initials: J
authors.lastname: Wang
mesh_terms: D015374:Biosensing Techniques; D002460:Cell Line; D004950:Esterases / Q000737:chemistry / Q000302:isolation & purification*; D006801:Humans; D057446:Hydrodynamics; D046210:Microfluidic Analytical Techniques; D011312:Pressure; D059010:Single-Cell Analysis
chemical_list: D004950:Esterases
keywords: Esterase heterogeneity; Microfluidics; Multiplex single-cell array; Pneumatic microvalve