Time Sequential Single-Cell Patterning with High Efficiency and High Density

Single-cell capture plays an important role in single-cell manipulation and analysis. This paper presents a microfluidic device for deterministic single-cell trapping based on the hydrodynamic trapping mechanism. The device is composed of an S-shaped loop channel and thousands of aligned trap units. This arrayed structure enables each row of the device to be treated equally and independently, as it has row periodicity. A theoretical model was established and a simulation was conducted to optimize the key geometric parameters, and the performance was evaluated by conducting experiments on MCF-7 and Jurkat cells. The results showed improvements in single-cell trapping ability, including loading efficiency, capture speed, and the density of the patterned cells. The optimized device can achieve a capture efficiency of up to 100% and single-cell capture efficiency of up to 95%. This device offers 200 trap units in an area of 1 mm2, which enables 100 single cells to be observed simultaneously using a microscope with a 20× objective lens. One thousand cells can be trapped sequentially within 2 min; this is faster than the values obtained with previously reported devices. Furthermore, the cells can also be recovered by reversely infusing solutions. The structure can be easily extended to a large scale, and a patterned array with 32,000 trap sites was accomplished on a single chip. This device can be a powerful tool for high-throughput single-cell analysis, cell heterogeneity investigation, and drug screening.

[1]  Kai Zhang,et al.  Block-Cell-Printing for live single-cell printing , 2014, Proceedings of the National Academy of Sciences.

[2]  Doryaneh Ahmadpour,et al.  Hydrodynamic Cell Trapping for High Throughput Single-Cell Applications , 2013, Micromachines.

[3]  Liang Huang,et al.  A fluidic circuit based, high-efficiency and large-scale single cell trap. , 2016, Lab on a chip.

[4]  G. Grau,et al.  Single-cell clones of liver cancer stem cells have the potential of differentiating into different types of tumor cells , 2013, Cell Death and Disease.

[5]  Xiaojing Zhong,et al.  On-Chip Studies of Magnetic Stimulation Effect on Single Neural Cell Viability and Proliferation on Glass and Nanoporous Surfaces. , 2018, ACS applied materials & interfaces.

[6]  Pierre Cosson,et al.  A microfluidic cell-trapping device for single-cell tracking of host-microbe interactions. , 2016, Lab on a chip.

[7]  D. Janasek,et al.  A microfluidic array with cellular valving for single cell co-culture. , 2011, Lab on a chip.

[8]  Ian Schneider,et al.  High Throughput Studies of Cell Migration in 3D Microtissues Fabricated by a Droplet Microfluidic Chip , 2016, Micromachines.

[9]  Jun Wang,et al.  Micropatterning of single cell arrays using the PEG-Silane and Biotin–(Strept)Avidin System with photolithography and chemical vapor deposition , 2013 .

[10]  Kumiko Miyajima,et al.  Separation and Analysis of Adherent and Non-Adherent Cancer Cells Using a Single-Cell Microarray Chip , 2017, Sensors.

[11]  D Jin,et al.  A microfluidic device enabling high-efficiency single cell trapping. , 2015, Biomicrofluidics.

[12]  A. Folch,et al.  Large-scale single-cell trapping and imaging using microwell arrays. , 2005, Analytical chemistry.

[13]  J. Voldman Electrical forces for microscale cell manipulation. , 2006, Annual review of biomedical engineering.

[14]  Long Que,et al.  Microtissue size and cell-cell communication modulate cell migration in arrayed 3D collagen gels , 2018, Biomedical microdevices.

[15]  Yiqiong Zhao,et al.  Using polarization-shaped optical vortex traps for single-cell nanosurgery. , 2007, Nano letters.

[16]  Sean C. Bendall,et al.  viSNE enables visualization of high dimensional single-cell data and reveals phenotypic heterogeneity of leukemia , 2013, Nature Biotechnology.

[17]  S. Vanapalli,et al.  Behavior of a train of droplets in a fluidic network with hydrodynamic traps. , 2010, Biomicrofluidics.

[18]  Dahai Ren,et al.  Multiplexed Analysis for Anti-Epidermal Growth Factor Receptor Tumor Cell Growth Inhibition Based on Quantum Dot Probes. , 2016, Analytical chemistry.

[19]  Gilbert Reyne,et al.  Diamagnetically trapped arrays of living cells above micromagnets. , 2011, Lab on a chip.

[20]  S. Digumarthy,et al.  Isolation of rare circulating tumour cells in cancer patients by microchip technology , 2007, Nature.

[21]  Ren Da Micropatterning and Its Applications in Biomedical Research , 2012 .

[22]  Zheng You,et al.  Multiplexed living cells stained with quantum dot bioprobes for multiplexed detection of single-cell array , 2013, Journal of biomedical optics.

[23]  B. W. Webb,et al.  Characterization of frictional pressure drop for liquid flows through microchannels , 2002 .

[24]  S. Lindström,et al.  Miniaturization of biological assays -- overview on microwell devices for single-cell analyses. , 2011, Biochimica et biophysica acta.

[25]  S. Garimella,et al.  Investigation of Liquid Flow in Microchannels , 2002 .

[26]  David J. Collins,et al.  Two-dimensional single-cell patterning with one cell per well driven by surface acoustic waves , 2015, Nature Communications.

[27]  Shoji Takeuchi,et al.  A trap-and-release integrated microfluidic system for dynamic microarray applications , 2007, Proceedings of the National Academy of Sciences.

[28]  Yiqiu Xia,et al.  Micropatterning and Its Applications in Biomedical Research*: Micropatterning and Its Applications in Biomedical Research* , 2012 .

[29]  Pilnam Kim,et al.  Microdroplet-based cell culture models and their application , 2016, BioChip Journal.

[30]  D. Kell,et al.  Flow cytometry and cell sorting of heterogeneous microbial populations: the importance of single-cell analyses. , 1996, Microbiological reviews.

[31]  Francesca Nason,et al.  Geometrical effects in microfluidic-based microarrays for rapid, efficient single-cell capture of mammalian stem cells and plant cells. , 2012, Biomicrofluidics.

[32]  A. Valero,et al.  Optimization of microfluidic single cell trapping for long-term on-chip culture. , 2010, Lab on a chip.

[33]  J. Voldman,et al.  A scalable addressable positive-dielectrophoretic cell-sorting array. , 2005, Analytical chemistry.

[34]  Luke P. Lee,et al.  Single-cell level co-culture platform for intercellular communication. , 2012, Integrative biology : quantitative biosciences from nano to macro.

[35]  Helene Andersson-Svahn,et al.  Overview of single-cell analyses: microdevices and applications. , 2010, Lab on a chip.

[36]  E. Mufson,et al.  Single cell gene expression profiling in Alzheimer’s disease , 2006, NeuroRX.