Hydrodynamic Cell Trapping for High Throughput Single-Cell Applications

The possibility to conduct complete cell assays under a precisely controlled environment while consuming minor amounts of chemicals and precious drugs have made microfluidics an interesting candidate for quantitative single-cell studies. Here, we present an application-specific microfluidic device, cellcomb, capable of conducting high-throughput single-cell experiments. The system employs pure hydrodynamic forces for easy cell trapping and is readily fabricated in polydimethylsiloxane (PDMS) using soft lithography techniques. The cell-trapping array consists of V-shaped pockets designed to accommodate up to six Saccharomyces cerevisiae (yeast cells) with the average diameter of 4 μm. We used this platform to monitor the impact of flow rate modulation on the arsenite (As(III)) uptake in yeast. Redistribution of a green fluorescent protein (GFP)-tagged version of the heat shock protein Hsp104 was followed over time as read out. Results showed a clear reverse correlation between the arsenite uptake and three different adjusted low = 25 nL min−1, moderate = 50 nL min−1, and high = 100 nL min−1 flow rates. We consider the presented device as the first building block of a future integrated application-specific cell-trapping array that can be used to conduct complete single cell experiments on different cell types.

[1]  Roland Zengerle,et al.  Microfluidic platforms for lab-on-a-chip applications. , 2007, Lab on a chip.

[2]  Mattias Goksör,et al.  Optical manipulation and microfluidics for studies of single cell dynamics , 2007 .

[3]  P. Swain,et al.  Stochastic Gene Expression in a Single Cell , 2002, Science.

[4]  Kerstin Ramser,et al.  Optical manipulation for single‐cell studies , 2010, Journal of biophotonics.

[5]  Luke P. Lee,et al.  Microfluidics-based systems biology. , 2006, Molecular bioSystems.

[6]  J. Y. Lim,et al.  Cell sensing and response to micro- and nanostructured surfaces produced by chemical and topographic patterning. , 2007, Tissue engineering.

[7]  Albert van den Berg,et al.  Single cells or large populations? , 2007, Lab on a chip.

[8]  Luke P. Lee,et al.  Dynamic single cell culture array. , 2006, Lab on a chip.

[9]  Catherine A. Rivet,et al.  Imaging single-cell signaling dynamics with a deterministic high-density single-cell trap array. , 2011, Analytical chemistry.

[10]  Markus J. Tamás,et al.  The glycerol channel Fps1p mediates the uptake of arsenite and antimonite in Saccharomyces cerevisiae , 2001, Molecular microbiology.

[11]  T. Nyström,et al.  A microfluidic system in combination with optical tweezers for analyzing rapid and reversible cytological alterations in single cells upon environmental changes. , 2007, Lab on a chip.

[12]  Robert Wysocki,et al.  The yeast aquaglyceroporin Fps1p is a bidirectional arsenite channel , 2010, FEBS letters.

[13]  Fabian Rudolf,et al.  Microfluidic single-cell cultivation chip with controllable immobilization and selective release of yeast cells. , 2012, Lab on a chip.

[14]  Mattias Goksör,et al.  CellStress - open source image analysis program for single-cell analysis , 2010, NanoScience + Engineering.

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

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

[17]  T. Elston,et al.  Stochasticity in gene expression: from theories to phenotypes , 2005, Nature Reviews Genetics.

[18]  Mattias Goksör,et al.  Acquisition of single cell data in an optical microscope , 2008 .

[19]  S. Quake,et al.  Microfluidics: Fluid physics at the nanoliter scale , 2005 .

[20]  Nam-Trung Nguyen,et al.  Oxygen plasma treatment for reducing hydrophobicity of a sealed polydimethylsiloxane microchannel. , 2010, Biomicrofluidics.

[21]  Albert van den Berg,et al.  Parallel single‐cell analysis microfluidic platform , 2011, Electrophoresis.

[22]  A. van Oudenaarden,et al.  Quantitative time-lapse fluorescence microscopy in single cells. , 2009, Annual review of cell and developmental biology.

[23]  Hiroyuki Kishi,et al.  Single-cell microarray for analyzing cellular response. , 2005, Analytical chemistry.

[24]  Stefan Hohmann,et al.  A Short Regulatory Domain Restricts Glycerol Transport through Yeast Fps1p* , 2003, The Journal of Biological Chemistry.

[25]  Clara Navarrete,et al.  Arsenite interferes with protein folding and triggers formation of protein aggregates in yeast , 2012, Journal of Cell Science.

[26]  Chun-Ping Jen,et al.  Single-Cell Chemical Lysis on Microfluidic Chips with Arrays of Microwells , 2011, Sensors.

[27]  Frederick F Becker,et al.  Microsample preparation by dielectrophoresis: isolation of malaria. , 2002, Lab on a chip.

[28]  Mehmet Toner,et al.  Cell handling using microstructured membranes. , 2006, Lab on a chip.

[29]  D. Figeys,et al.  Lab-on-a-chip: a revolution in biological and medical sciences , 2000, Analytical chemistry.

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

[31]  Aaron R Wheeler,et al.  Microfluidic device for single-cell analysis. , 2003, Analytical chemistry.

[32]  A. Ashkin,et al.  Optical trapping and manipulation of single cells using infrared laser beams , 1987, Nature.

[33]  Zachary R. Gagnon,et al.  Cellular dielectrophoresis: Applications to the characterization, manipulation, separation and patterning of cells , 2011, Electrophoresis.

[34]  Mattias Goksör,et al.  A microfluidic device for reversible environmental changes around single cells using optical tweezers for cell selection and positioning. , 2010, Lab on a chip.

[35]  Stephen A. Bustin,et al.  Real-Time PCR , 2005 .

[36]  W. N. Burnette,et al.  "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. , 1981, Analytical biochemistry.

[37]  M. Textor,et al.  Surface engineering approaches to micropattern surfaces for cell-based assays. , 2006, Biomaterials.

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