Recent advances in single-cell analysis using capillary electrophoresis and microfluidic devices.

Cells are the fundamental unit of life, and studies on cell contribute to reveal the mystery of life. However, since variability exists between individual cells even in the same kind of cells, increased emphasis has been put on the analysis of individual cells for getting better understanding on the organism functions. During the past two decades, various techniques have been developed for single-cell analysis. Capillary electrophoresis is an excellent technique for identifying and quantifying the contents of single cells. The microfluidic devices afford a versatile platform for single-cell analysis owing to their unique characteristics. This article provides a review on recent advances in single-cell analysis using capillary electrophoresis and microfluidic devices; focus areas to be covered include sampling techniques, detection methods and main applications in capillary electrophoresis, and cell culture, cell manipulation, chemical cytometry and cellular physiology on microfluidic devices.

[1]  B. Chung,et al.  Human neural stem cell growth and differentiation in a gradient-generating microfluidic device. , 2005, Lab on a chip.

[2]  C. Gillmor,et al.  Multiple sampling in single-cell enzyme assays using CE-laser-induced fluorescence to monitor reaction progress. , 2005, Analytical chemistry.

[3]  J. Sweedler,et al.  A multichannel native fluorescence detection system for capillary electrophoretic analysis of neurotransmitters in single neurons , 2006, Analytical and bioanalytical chemistry.

[4]  Bo Huang,et al.  Counting Low-Copy Number Proteins in a Single Cell , 2007, Science.

[5]  Wilhelm Pfleging,et al.  A chip-based platform for the in vitro generation of tissues in three-dimensional organization. , 2007, Lab on a chip.

[6]  Alexandra Ros,et al.  Bioanalysis in structured microfluidic systems , 2006, Electrophoresis.

[7]  S J Lillard,et al.  Continuous cell introduction for the analysis of individual cells by capillary electrophoresis. , 2001, Analytical chemistry.

[8]  Stephen R. Quake,et al.  Microfluidic Digital PCR Enables Multigene Analysis of Individual Environmental Bacteria , 2006, Science.

[9]  Wei-Hua Huang,et al.  Recent developments in single-cell analysis , 2004 .

[10]  Christian Amatore,et al.  Electrochemical detection in a microfluidic device of oxidative stress generated by macrophage cells. , 2007, Lab on a chip.

[11]  Alexandra Ros,et al.  Single cell manipulation, analytics, and label‐free protein detection in microfluidic devices for systems nanobiology , 2005, Electrophoresis.

[12]  Joel Voldman,et al.  Cell patterning chip for controlling the stem cell microenvironment. , 2007, Biomaterials.

[13]  Elinore M Mercer,et al.  Microfluidic sorting of mammalian cells by optical force switching , 2005, Nature Biotechnology.

[14]  T. Laurell,et al.  Free flow acoustophoresis: microfluidic-based mode of particle and cell separation. , 2007, Analytical chemistry.

[15]  Stephen R Quake,et al.  Microfluidic single-cell mRNA isolation and analysis. , 2006, Analytical chemistry.

[16]  Hua Xiao,et al.  Immunoassay of P-glycoprotein on single cell by capillary electrophoresis with laser induced fluorescence detection , 2006 .

[17]  D. J. Harrison,et al.  Immunomagnetic T cell capture from blood for PCR analysis using microfluidic systems. , 2004, Lab on a chip.

[18]  Thomas Laurell,et al.  Noninvasive acoustic cell trapping in a microfluidic perfusion system for online bioassays. , 2007, Analytical chemistry.

[19]  Robert T Kennedy,et al.  Perfusion and chemical monitoring of living cells on a microfluidic chip. , 2005, Lab on a chip.

[20]  G. Whitesides,et al.  Patterning proteins and cells using soft lithography. , 1999, Biomaterials.

[21]  Yanming Liu,et al.  Capillary electrophoresis immunoassay chemiluminescence detection of zeptomoles of bone morphogenic protein-2 in rat vascular smooth muscle cells. , 2004, Analytical chemistry.

[22]  Jicun Ren,et al.  Coupling chemiluminescence with capillary electrophoresis to analyze single human red blood cells. , 2007, Analytica chimica acta.

[23]  Luke P. Lee,et al.  Mammalian electrophysiology on a microfluidic platform. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[24]  J. Sweedler,et al.  Direct single cell determination of nitric oxide synthase related metabolites in identified nitrergic neurons. , 2005, Journal of inorganic biochemistry.

[25]  Luke P. Lee,et al.  Microfluidic application-specific integrated device for monitoring direct cell-cell communication via gap junctions between individual cell pairs , 2005 .

[26]  Measuring nitric oxide in single neurons by capillary electrophoresis with laser-induced fluorescence: use of ascorbate oxidase in diaminofluorescein measurements. , 2006, Analytical chemistry.

[27]  Wei-Hua Huang,et al.  Transport, location, and quantal release monitoring of single cells on a microfluidic device. , 2004, Analytical chemistry.

[28]  C. Cotman,et al.  A microfluidic culture platform for CNS axonal injury, regeneration and transport , 2005, Nature Methods.

[29]  W. Jin,et al.  Determination of different forms of human interferon‐γ in single natural killer cells by capillary electrophoresis with on‐capillary immunoreaction and laser‐induced fluorescence detection , 2004, Electrophoresis.

[30]  Tejal A Desai,et al.  Layer-by-layer microfluidics for biomimetic three-dimensional structures. , 2004, Biomaterials.

[31]  Xuemei Sun,et al.  Measurement of alkaline phosphatase isoenzymes in individual mouse bone marrow fibroblast cells based on capillary electrophoresis with on‐capillary enzyme‐catalyzed reaction and electrochemical detection , 2004, Electrophoresis.

[32]  H. Kleinman,et al.  Use of extracellular matrix components for cell culture. , 1987, Analytical biochemistry.

[33]  Michael A. Rodriguez,et al.  Capillary electrophoretic method for the detection of bacterial contamination. , 2006, Analytical chemistry.

[34]  Robert M Johann,et al.  Cell trapping in microfluidic chips , 2006, Analytical and bioanalytical chemistry.

[35]  Mengsu Yang,et al.  Microfluidics technology for manipulation and analysis of biological cells , 2006 .

[36]  Cheng-Wey Wei,et al.  Elucidating in vitro cell-cell interaction using a microfluidic coculture system , 2006, Biomedical microdevices.

[37]  Alexandra Ros,et al.  Improved native UV laser induced fluorescence detection for single cell analysis in poly(dimethylsiloxane) microfluidic devices. , 2006, Journal of chromatography. A.

[38]  Luke P. Lee,et al.  An artificial liver sinusoid with a microfluidic endothelial-like barrier for primary hepatocyte culture. , 2007, Biotechnology and bioengineering.

[39]  Godfrey L. Smith,et al.  Microfluidic systems to examine intercellular coupling of pairs of cardiac myocytes. , 2007, Lab on a chip.

[40]  J. Jorgenson,et al.  Microcolumn separations and the analysis of single cells. , 1989, Science.

[41]  Shuichi Takayama,et al.  Computerized microfluidic cell culture using elastomeric channels and Braille displays. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[42]  A. Ewing,et al.  Capillary zone electrophoresis with electrochemical detection in 12.7 microns diameter columns. , 1988, Analytical chemistry.

[43]  C. Culbertson,et al.  Chemical analysis of single mammalian cells with microfluidics. Strategies for culturing, sorting, trapping, and lysing cells and separating their contents on chips. , 2007, Analytical chemistry.

[44]  Hanry Yu,et al.  A novel 3D mammalian cell perfusion-culture system in microfluidic channels. , 2007, Lab on a chip.

[45]  D. Armstrong,et al.  Single-cell detection: test of microbial contamination using capillary electrophoresis. , 2007, Analytical chemistry.

[46]  K. Jensen,et al.  Cells on chips , 2006, Nature.

[47]  D. Chiu,et al.  Selective encapsulation of single cells and subcellular organelles into picoliter- and femtoliter-volume droplets. , 2005, Analytical chemistry.

[48]  Xin Liu,et al.  Determination of single-cell gene expression in Arabidopsis by capillary electrophoresis with laser induced fluorescence detection. , 2004, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[49]  Albert Folch,et al.  Differentiation-on-a-chip: a microfluidic platform for long-term cell culture studies. , 2005, Lab on a chip.

[50]  Xue-feng Yin,et al.  Intracellular labeling method for chip-based capillary electrophoresis fluorimetric single cell analysis using liposomes. , 2006, Journal of chromatography. A.

[51]  E. Arriaga,et al.  Analysis of individual acidic organelles by capillary electrophoresis with laser-induced fluorescence detection facilitated by the endocytosis of fluorescently labeled microspheres. , 2003, Analytical chemistry.

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

[53]  Gregory T. Roman,et al.  Single-cell manipulation and analysis using microfluidic devices , 2006, Analytical and bioanalytical chemistry.

[54]  R. M. Westervelt,et al.  Dielectrophoresis tweezers for single cell manipulation , 2006, Biomedical microdevices.

[55]  E. P. Furlani,et al.  Magnetophoretic separation of blood cells at the microscale , 2006, physics/0612005.

[56]  Nancy L Allbritton,et al.  CRITICAL REVIEW www.rsc.org/loc | Lab on a Chip Analysis of single mammalian cells on-chip , 2006 .

[57]  D. Chiu,et al.  Thermoelectric manipulation of aqueous droplets in microfluidic devices. , 2007, Analytical chemistry.

[58]  Weihua Huang,et al.  Carbon fiber nanoelectrodes applied to microchip electrophoresis amperometric detection of neurotransmitter dopamine in rat pheochromocytoma (PC12) cells , 2007, Electrophoresis.

[59]  V. Vandelinder,et al.  Perfusion in microfluidic cross-flow: separation of white blood cells from whole blood and exchange of medium in a continuous flow. , 2007, Analytical chemistry.

[60]  Zhao-Lun Fang,et al.  Integration of single cell injection, cell lysis, separation and detection of intracellular constituents on a microfluidic chip. , 2004, Lab on a chip.

[61]  Michael G. Roper,et al.  Microfluidic chip for continuous monitoring of hormone secretion from live cells using an electrophoresis-based immunoassay. , 2003, Analytical chemistry.

[62]  Chen Yu,et al.  Magnetic-based microfluidic platform for biomolecular separation , 2006, Biomedical microdevices.

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

[64]  N. Chronis,et al.  Electrothermally activated SU-8 microgripper for single cell manipulation in solution , 2005, Journal of Microelectromechanical Systems.

[65]  N. Dovichi,et al.  Single-cell protein analysis of a single mouse embryo by two-dimensional capillary electrophoresis. , 2006, Journal of chromatography. A.

[66]  Xingyu Jiang,et al.  A method for patterning multiple types of cells by using electrochemical desorption of self-assembled monolayers within microfluidic channels. , 2007, Angewandte Chemie.

[67]  Hanry Yu,et al.  A practical guide to microfluidic perfusion culture of adherent mammalian cells. , 2007, Lab on a chip.

[68]  Ming C. Wu,et al.  Massively parallel manipulation of single cells and microparticles using optical images , 2005, Nature.

[69]  Chia-Chi Ho,et al.  Biocompatible micropatterning of two different cell types. , 2005, Journal of the American Chemical Society.

[70]  Godfrey L. Smith,et al.  Metabolic monitoring of the electrically stimulated single heart cell within a microfluidic platform. , 2006, Lab on a chip.

[71]  E. Arriaga,et al.  Analysis of mitochondria isolated from single cells , 2006, Analytical and bioanalytical chemistry.

[72]  Mark Bachman,et al.  Micropallet arrays for the separation of single, adherent cells. , 2007, Analytical chemistry.

[73]  Cheng-Hsien Liu,et al.  Rapid heterogeneous liver-cell on-chip patterning via the enhanced field-induced dielectrophoresis trap. , 2006, Lab on a chip.

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

[75]  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.

[76]  Mehmet Toner,et al.  Size-based microfluidic enrichment of neonatal rat cardiac cell populations , 2006, Biomedical microdevices.

[77]  Luke P. Lee,et al.  Open-access microfluidic patch-clamp array with raised lateral cell trapping sites. , 2006, Lab on a chip.

[78]  S. Lillard,et al.  A qualitative look at multiplex gene expression of single cells using capillary electrophoresis , 2005, Electrophoresis.

[79]  Weihua Huang,et al.  Monitoring dopamine release from single living vesicles with nanoelectrodes. , 2005, Journal of the American Chemical Society.

[80]  Andrew G Ewing,et al.  Analysis of Mammalian Cell Cytoplasm with Electrophoresis in Nanometer Inner Diameter Capillaries. , 2005, Electroanalysis.

[81]  W. Jin,et al.  Single-cell analysis by intracellular immuno-reaction and capillary electrophoresis with laser-induced fluorescence detection. , 2006, Journal of chromatography. A.

[82]  Dorian Liepmann,et al.  Biomimetic technique for adhesion-based collection and separation of cells in a microfluidic channel. , 2005, Lab on a chip.

[83]  J. Sweedler,et al.  Measuring D-amino acid-containing neuropeptides with capillary electrophoresis. , 2005, The Analyst.

[84]  Robert Johann,et al.  Gentle cell trapping and release on a microfluidic chip by in situ alginate hydrogel formation. , 2005, Lab on a chip.

[85]  David A Michels,et al.  Capillary sieving electrophoresis/micellar electrokinetic capillary chromatography for two-dimensional protein fingerprinting of single mammalian cells. , 2004, Analytical chemistry.

[86]  Dino Di Carlo,et al.  Dynamic single-cell analysis for quantitative biology. , 2006, Analytical chemistry.

[87]  N. Dovichi,et al.  Metabolic cytometry. Glycosphingolipid metabolism in single cells. , 2007, Analytical chemistry.

[88]  R. Wightman Probing Cellular Chemistry in Biological Systems with Microelectrodes , 2006, Science.

[89]  J. Sweedler,et al.  Subcellular analysis of D-aspartate. , 2005, Analytical chemistry.

[90]  Barry R Lutz,et al.  Hydrodynamic tweezers: 1. Noncontact trapping of single cells using steady streaming microeddies. , 2006, Analytical chemistry.

[91]  Jieke Cheng,et al.  Determination of neurotransmitters in PC 12 cells by microchip electrophoresis with fluorescence detection , 2007, Electrophoresis.

[92]  R. Tompkins,et al.  Effect of flow and surface conditions on human lymphocyte isolation using microfluidic chambers. , 2004, Langmuir : the ACS journal of surfaces and colloids.

[93]  Mihrimah Ozkan,et al.  Manipulation of microspheres and biological cells with multiple agile VCSEL traps , 2006 .

[94]  Donald E Ingber,et al.  Combined microfluidic-micromagnetic separation of living cells in continuous flow , 2006, Biomedical microdevices.

[95]  John F. Dishinger,et al.  Serial immunoassays in parallel on a microfluidic chip for monitoring hormone secretion from living cells. , 2007, Analytical chemistry.

[96]  A. Ewing,et al.  Electrically assisted sampling across membranes with electrophoresis in nanometer inner diameter capillaries. , 2005, Analytical chemistry.

[97]  R. Zare,et al.  Chemical cytometry on a picoliter-scale integrated microfluidic chip. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[98]  Luke P. Lee,et al.  Continuous perfusion microfluidic cell culture array for high-throughput cell-based assays. , 2005, Biotechnology and bioengineering.

[99]  Luke P. Lee,et al.  A novel high aspect ratio microfluidic design to provide a stable and uniform microenvironment for cell growth in a high throughput mammalian cell culture array. , 2005, Lab on a chip.

[100]  M. Eggertson,et al.  Instrumentation for medium-throughput two-dimensional capillary electrophoresis with laser-induced fluorescence detection. , 2007, Analytical chemistry.

[101]  Brian P Helmke,et al.  Designing a nano-interface in a microfluidic chip to probe living cells: challenges and perspectives. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

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

[103]  N. Dovichi,et al.  Detection of green fluorescent protein in a single bacterium by capillary electrophoresis with laser-induced fluorescence. , 2007, Analytical chemistry.

[104]  Robert Langer,et al.  Hyaluronic acid hydrogel for controlled self-renewal and differentiation of human embryonic stem cells , 2007, Proceedings of the National Academy of Sciences.

[105]  Andreas Offenhäusser,et al.  Micropatterned Substrates for the Growth of Functional Neuronal Networks of Defined Geometry , 2003, Biotechnology progress.

[106]  Michael F. Santillo,et al.  Recent advances in capillary electrophoretic analysis of individual cells , 2006, Analytical and bioanalytical chemistry.