Current Trends of Microfluidic Single-Cell Technologies

The investigation of human disease mechanisms is difficult due to the heterogeneity in gene expression and the physiological state of cells in a given population. In comparison to bulk cell measurements, single-cell measurement technologies can provide a better understanding of the interactions among molecules, organelles, cells, and the microenvironment, which can aid in the development of therapeutics and diagnostic tools. In recent years, single-cell technologies have become increasingly robust and accessible, although limitations exist. In this review, we describe the recent advances in single-cell technologies and their applications in single-cell manipulation, diagnosis, and therapeutics development.

[1]  R N Zare,et al.  Nanoengineered structures for holding and manipulating liposomes and cells. , 2001, Analytical chemistry.

[2]  J. Sweedler,et al.  Single-cell MALDI: a new tool for direct peptide profiling. , 2000, Trends in biotechnology.

[3]  Sangjin Park,et al.  Carbon nanosyringe array as a platform for intracellular delivery. , 2009, Nano letters.

[4]  A. Schmid,et al.  Single-cell analysis in biotechnology, systems biology, and biocatalysis. , 2012, Annual review of chemical and biomolecular engineering.

[5]  Chad A. Shaw,et al.  Evidence for feasibility of fetal trophoblastic cell‐based noninvasive prenatal testing† , 2016, Prenatal diagnosis.

[6]  Tapobrata Panda,et al.  Electroporation: basic principles, practical considerations and applications in molecular biology , 1997 .

[7]  Romain Quidant,et al.  Plasmon nano-optical tweezers , 2011 .

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

[9]  Lu Wen,et al.  Single-cell triple omics sequencing reveals genetic, epigenetic, and transcriptomic heterogeneity in hepatocellular carcinomas , 2016, Cell Research.

[10]  Michael R Hamblin,et al.  Physical energy for drug delivery; poration, concentration and activation. , 2014, Advanced drug delivery reviews.

[11]  James C. Weaver,et al.  High-throughput Nuclear Delivery and Rapid Expression of DNA via Mechanical and Electrical Cell-Membrane Disruption , 2017, Nature Biomedical Engineering.

[12]  E. Yoon,et al.  Scaling and automation of a high-throughput single-cell-derived tumor sphere assay chip. , 2016, Lab on a chip.

[13]  O Orwar,et al.  Altering the biochemical state of individual cultured cells and organelles with ultramicroelectrodes. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[14]  Peter Kuhn,et al.  Multiplex protein detection on circulating tumor cells from liquid biopsies using imaging mass cytometry. , 2018, Convergent science physical oncology.

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

[16]  John A. Cole,et al.  Heterogeneity in protein expression induces metabolic variability in a modeled Escherichia coli population , 2013, Proceedings of the National Academy of Sciences.

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

[18]  Norihisa Miki,et al.  Round-tip dielectrophoresis-based tweezers for single micro-object manipulation. , 2013, Biosensors & bioelectronics.

[19]  Fan-Gang Tseng,et al.  Recent Trends on Micro/Nanofluidic Single Cell Electroporation , 2013, Micromachines.

[20]  William H. Grover,et al.  Using buoyant mass to measure the growth of single cells , 2010, Nature Methods.

[21]  Joshua M. Kunken,et al.  Fluid biopsy in patients with metastatic prostate, pancreatic and breast cancers , 2012, Physical biology.

[22]  O Orwar,et al.  Characterization of single-cell electroporation by using patch-clamp and fluorescence microscopy. , 2000, Biophysical journal.

[23]  I. Weissman,et al.  Stem cells, cancer, and cancer stem cells , 2001, Nature.

[24]  Yi Liu,et al.  Enhanced and Differential Capture of Circulating Tumor Cells from Lung Cancer Patients by Microfluidic Assays Using Aptamer Cocktail. , 2016, Small.

[25]  Sungmin Son,et al.  Direct observation of mammalian cell growth and size regulation , 2012, Nature Methods.

[26]  H. Beutel,et al.  Fast and precise positioning of single cells on planar electrode substrates , 1999, IEEE Engineering in Medicine and Biology Magazine.

[27]  Donald Wlodkowic,et al.  Microfluidic single-cell array cytometry for the analysis of tumor apoptosis. , 2009, Analytical chemistry.

[28]  Richard A Mathies,et al.  Microfluidic device for electric field-driven single-cell capture and activation. , 2005, Analytical chemistry.

[29]  Sreeurpa Ray,et al.  The Cell: A Molecular Approach , 1996 .

[30]  M. Casanova,et al.  Altered expression of Alzheimer's disease-related genes in the cerebellum of autistic patients: a model for disrupted brain connectome and therapy , 2014, Cell Death and Disease.

[31]  Michael A. Teitell,et al.  Photothermal nanoblade for patterned cell membrane cutting , 2010, Optics express.

[32]  Min-Sung Kim,et al.  Generating mammalian stable cell lines by electroporation. , 2013, Methods in enzymology.

[33]  Keegan D. Korthauer,et al.  A statistical approach for identifying differential distributions in single-cell RNA-seq experiments , 2016, Genome Biology.

[34]  J. E. Duarte,et al.  Bacterial handling under the influence of non-uniform electric fields: dielectrophoretic and electrohydrodynamic effects. , 2008, Anais da Academia Brasileira de Ciencias.

[35]  Pallavi Shinde,et al.  Mechanoporation: Toward Single Cell Approaches , 2021, Handbook of Single-Cell Technologies.

[36]  R Pool,et al.  Trapping with optical tweezers. , 1988, Science.

[37]  Stuart J. Williams,et al.  Electrokinetic patterning of colloidal particles with optical landscapes. , 2008, Lab on a chip.

[38]  A. Souka,et al.  Identification of nucleated red blood cells in maternal circulation: A second step in screening for fetal aneuploidies and pregnancy complications , 2007, Prenatal diagnosis.

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

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

[41]  Sarvesh Sukhatme,et al.  Digital Microfluidics: Techniques, Their Applications and Advantages , 2013 .

[42]  Duo Zhang,et al.  Enrichment of selective miRNAs in exosomes and delivery of exosomal miRNAs in vitro and in vivo. , 2017, American journal of physiology. Lung cellular and molecular physiology.

[43]  Fan-Gang Tseng,et al.  Essentials of Single-Cell Analysis Concepts, Applications and Future Prospects , 2016 .

[44]  Duncan Casey,et al.  Optical Tools for Single-Cell Manipulation and Analysis , 2016 .

[45]  A. Theberge,et al.  Microdroplets in microfluidics: an evolving platform for discoveries in chemistry and biology. , 2010, Angewandte Chemie.

[46]  M. Pang,et al.  A high yield of fetal nucleated red blood cells isolated using optimal osmolality and a double‐density gradient system , 2007, Prenatal diagnosis.

[47]  Nassir Navab,et al.  A BaSiC tool for background and shading correction of optical microscopy images , 2017, Nature Communications.

[48]  Yu Sun,et al.  Microfluidic approaches for cancer cell detection, characterization, and separation. , 2012, Lab on a chip.

[49]  Hugo Germain,et al.  Droplet Digital PCR versus qPCR for gene expression analysis with low abundant targets: from variable nonsense to publication quality data , 2017, Scientific Reports.

[50]  Andrey Tovchigrechko,et al.  Recombinant human B cell repertoires enable screening for rare, specific, and natively paired antibodies , 2018, Communications Biology.

[51]  A. Beaudet,et al.  Using fetal cells for prenatal diagnosis: History and recent progress , 2016, American journal of medical genetics. Part C, Seminars in medical genetics.

[52]  Jan Scrimgeour,et al.  Live cell lithography: using optical tweezers to create synthetic tissue. , 2008, Lab on a chip.

[53]  Alfred Cuschieri,et al.  Magnetoporation and magnetolysis of cancer cells via carbon nanotubes induced by rotating magnetic fields. , 2012, Nano letters.

[54]  Shuang Hou,et al.  Detecting ALK-rearrangement of CTC enriched by nanovelcro chip in advanced NSCLC patients. , 2016, Oncotarget.

[55]  G. Whitesides.  The origins and the future of microfluidics , 2006, Nature.

[56]  J. Voldman,et al.  Dielectrophoretic registration of living cells to a microelectrode array. , 2004, Biosensors & bioelectronics.

[57]  Aparajita Dutta,et al.  Validation of automatic scanning of microscope slides in recovering rare cellular events: application for detection of fetal cells in maternal blood , 2014, Prenatal diagnosis.

[58]  Jorge Nocedal,et al.  On the limited memory BFGS method for large scale optimization , 1989, Math. Program..

[59]  N. Manaresi,et al.  A CMOS chip for individual cell manipulation and detection , 2003, 2003 IEEE International Solid-State Circuits Conference, 2003. Digest of Technical Papers. ISSCC..

[60]  K. Greulich,et al.  Micromanipulation by laser microbeam and optical tweezers: from plant cells to single molecules , 2000, Journal of microscopy.

[61]  C. Bertozzi,et al.  Site-Specific Antibody–Drug Conjugates: The Nexus of Bioorthogonal Chemistry, Protein Engineering, and Drug Development , 2014, Bioconjugate chemistry.

[62]  R. Zengerle,et al.  Inkjet-like printing of single-cells. , 2011, Lab on a chip.

[63]  Hyoung Won Baac,et al.  Micro-ultrasonic cleaving of cell clusters by laser-generated focused ultrasound and its mechanisms. , 2013, Biomedical optics express.

[64]  P Sarder,et al.  Statistical Design of Position-Encoded Microsphere Arrays , 2011, IEEE Transactions on NanoBioscience.

[65]  N. Perrimon,et al.  Droplet microfluidic technology for single-cell high-throughput screening , 2009, Proceedings of the National Academy of Sciences.

[66]  Jason R Swedlow,et al.  Innovation in biological microscopy: Current status and future directions , 2012, BioEssays : news and reviews in molecular, cellular and developmental biology.

[67]  Ripudaman Singh,et al.  Genome‐wide copy number analysis on DNA from fetal cells isolated from the blood of pregnant women , 2016, Prenatal diagnosis.

[68]  Jordan R. Myers,et al.  Ultra-High Resolution 3D Imaging of Whole Cells , 2016, Cell.

[69]  Alexander van Oudenaarden,et al.  References and Notes Supporting Online Material Circadian Gating of the Cell Cycle Revealed in Single Cyanobacterial Cells , 2022 .

[70]  Antoni Ribas,et al.  Single-cell analysis tools for drug discovery and development , 2015, Nature Reviews Drug Discovery.

[71]  Stuart J. Williams,et al.  Characterization of 2D colloid aggregations created by optically induced electrohydrodynamics , 2015, Electrophoresis.

[72]  Feng Guo,et al.  Fetal nucleated red blood cell analysis for non-invasive prenatal diagnostics using a nanostructure microchip. , 2017, Journal of materials chemistry. B.

[73]  Kwang-Seok Yun,et al.  Single-cell manipulation on microfluidic chip by dielectrophoretic actuation and impedance detection , 2010 .

[74]  C. Vakoc,et al.  Targeting Transcription Factors in Cancer. , 2015, Trends in cancer.

[75]  Tony Fields,et al.  Novel Cell and Tissue Acquisition System (CTAS): Microdissection of Live and Frozen Brain Tissues , 2012, PloS one.

[76]  Mario Roederer,et al.  Single-cell technologies for monitoring immune systems , 2014, Nature Immunology.

[77]  Arye Nehorai,et al.  Optimization of microfluidic microsphere-trap arrays. , 2013, Biomicrofluidics.

[78]  Mohsen Soleimani,et al.  Synthesis of a functional metal-chelating polymer and steps toward quantitative mass cytometry bioassays. , 2010, Analytical chemistry.

[79]  Fumihito Arai,et al.  Screening of single Escherichia coli in a microchannel system by electric field and laser tweezers , 1998 .

[80]  Felix Naef,et al.  Circadian Gene Expression in Individual Fibroblasts Cell-Autonomous and Self-Sustained Oscillators Pass Time to Daughter Cells , 2004, Cell.

[81]  Menake E Piyasena,et al.  The intersection of flow cytometry with microfluidics and microfabrication. , 2014, Lab on a chip.

[82]  Fengchao Wang,et al.  A high throughput platform for stem cell-niche co-cultures and downstream gene expression analysis , 2015, Nature Cell Biology.

[83]  Francisco Feijó Delgado,et al.  High-throughput measurement of single-cell growth rates using serial microfluidic mass sensor arrays , 2016, Nature Biotechnology.

[84]  Buddy D Ratner,et al.  Cell sheet detachment affects the extracellular matrix: a surface science study comparing thermal liftoff, enzymatic, and mechanical methods. , 2005, Journal of biomedical materials research. Part A.

[85]  Shuichi Takayama,et al.  Simple Multi-level Microchannel Fabrication by Pseudo-Grayscale Backside Diffused Light Lithography. , 2013, RSC advances.

[86]  Ying Liu,et al.  Deficient brain insulin signalling pathway in Alzheimer's disease and diabetes , 2011, The Journal of pathology.

[87]  Fan-Gang Tseng,et al.  Micro/Nanofluidic Devices for Single Cells Analysis , 2014, Micromachines.

[88]  Shuichi Shoji,et al.  High-throughput single-cell manipulation system for a large number of target cells. , 2011, Biomicrofluidics.

[89]  Yunpeng Li,et al.  CIDRE: an illumination-correction method for optical microscopy , 2015, Nature Methods.

[90]  Aloke Kumar,et al.  Optoelectrokinetic Manipulation for Cell Analysis , 2016 .

[91]  Dexi Liu,et al.  Physical methods for gene transfer. , 2015, Advances in genetics.

[92]  James K. Mills,et al.  Cleavage-stage embryo rotation tracking and automated micropipette control: Towards automated single cell manipulation , 2016, 2016 IEEE/RSJ International Conference on Intelligent Robots and Systems (IROS).

[93]  Franziska Michor,et al.  In situ single cell analysis identifies heterogeneity for PIK3CA mutation and HER2 amplification in HER2+ breast cancer , 2015, Nature Genetics.

[94]  Woo-Jin Chang,et al.  Automated Dielectrophoretic Tweezers-Based Force Spectroscopy System in a Microfluidic Device , 2017, Sensors.

[95]  Keekyoung Kim,et al.  3D bioprinting for engineering complex tissues. , 2016, Biotechnology advances.

[96]  Chunbo Li,et al.  Tanshinone IIA reduces the risk of Alzheimer's disease by inhibiting iNOS, MMP‑2 and NF‑κBp65 transcription and translation in the temporal lobes of rat models of Alzheimer's disease. , 2014, Molecular medicine reports.

[97]  Subra Suresh,et al.  Three-dimensional manipulation of single cells using surface acoustic waves , 2016, Proceedings of the National Academy of Sciences.

[98]  Dong Sun,et al.  Enhanced cell sorting and manipulation with combined optical tweezer and microfluidic chip technologies. , 2011, Lab on a chip.

[99]  Sean C. Bendall,et al.  A deep profiler's guide to cytometry. , 2012, Trends in immunology.

[100]  Michael P. Sheetz,et al.  Cell Spreading and Lamellipodial Extension Rate Is Regulated by Membrane Tension , 2000, The Journal of cell biology.

[101]  Steven R. Head,et al.  Next-generation sequencing , 2010, Nature Reviews Drug Discovery.

[102]  Kwang-Seok Yun,et al.  Micro/Nanofluidic Device for Single-Cell-Based Assay , 2005, Biomedical microdevices.

[103]  Zhixiong Zhang,et al.  Microfluidics 3D gel-island chip for single cell isolation and lineage-dependent drug responses study. , 2016, Lab on a chip.

[104]  Michel Meunier,et al.  Visible and near infrared resonance plasmonic enhanced nanosecond laser optoporation of cancer cells , 2013, Biomedical optics express.

[105]  Itamar Simon,et al.  Lineage correlations of single cell division time as a probe of cell-cycle dynamics , 2015, Nature.

[106]  Meike T. Wortel,et al.  Lost in Transition: Start-Up of Glycolysis Yields Subpopulations of Nongrowing Cells , 2014, Science.

[107]  Kevin Braeckmans,et al.  Comparison of gold nanoparticle mediated photoporation: vapor nanobubbles outperform direct heating for delivering macromolecules in live cells. , 2014, ACS nano.

[108]  M. Fulwyler.  STATUS QUO IN FLOW-THROUGH CYTOMETRY , 1974, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[109]  George Whitesides.  The lab finally comes to the chip! , 2014, Lab on a chip.

[110]  Reinhard Schliebs,et al.  Down‐regulation of muscarinic acetylcholine receptor M2 adversely affects the expression of Alzheimer's disease‐relevant genes and proteins , 2005, Journal of neurochemistry.

[111]  Hiroyuki Kishi,et al.  A rapid and efficient single-cell manipulation method for screening antigen-specific antibody–secreting cells from human peripheral blood , 2009, Nature Medicine.

[112]  Péter Fürjes,et al.  Automated single cell isolation from suspension with computer vision , 2016, Scientific Reports.

[113]  Sang Heon Lee,et al.  High Density Individually Addressable Nanowire Arrays Record Intracellular Activity from Primary Rodent and Human Stem Cell Derived Neurons. , 2017, Nano letters.

[114]  Yi-Kuen Lee,et al.  Highly efficient capture of circulating tumor cells by using nanostructured silicon substrates with integrated chaotic micromixers. , 2011, Angewandte Chemie.

[115]  Andrew G Ewing,et al.  Hybrid capillary-microfluidic device for the separation, lysis, and electrochemical detection of vesicles. , 2009, Analytical chemistry.

[116]  Kevin Braeckmans,et al.  Laser-assisted photoporation: fundamentals, technological advances and applications , 2016 .

[117]  David G. Kirsch,et al.  Application of single-cell RNA sequencing in optimizing a combinatorial therapeutic strategy in metastatic renal cell carcinoma , 2016, Genome Biology.

[118]  Toshiyuki Takagi,et al.  On-demand killing of adherent cells on photo-acid-generating culture substrates. , 2013, Biotechnology and bioengineering.

[119]  Pallavi Shinde,et al.  Single-cell electroporation: current trends, applications and future prospects , 2018, Journal of Micromechanics and Microengineering.

[120]  Shuang Hou,et al.  Subclassification of prostate cancer circulating tumor cells by nuclear size reveals very small nuclear circulating tumor cells in patients with visceral metastases , 2015, Cancer.

[121]  Bernhard H Weigl,et al.  Microfluidic technologies in clinical diagnostics. , 2002, Clinica chimica acta; international journal of clinical chemistry.

[122]  Shawn M. Gillespie,et al.  Single-cell RNA-seq highlights intratumoral heterogeneity in primary glioblastoma , 2014, Science.

[123]  Catherine A. Blish,et al.  Application of Mass Cytometry (CyTOF) for Functional and Phenotypic Analysis of Natural Killer Cells. , 2016, Methods in molecular biology.

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

[125]  Richard Bruch,et al.  Multiplexed Point-of-Care Testing – xPOCT , 2017, Trends in biotechnology.

[126]  Robert J. Thomas,et al.  Practical guide to ICP-MS , 2003 .

[127]  J. Hoffmann,et al.  Reference values of fetal erythrocytes in maternal blood during pregnancy established using flow cytometry. , 2011, American journal of clinical pathology.

[128]  Sara Linse,et al.  A Microfluidic Platform for Real-Time Detection and Quantification of Protein-Ligand Interactions. , 2016, Biophysical journal.

[129]  R. Schmid,et al.  Liposome mediated gene transfer into the rat oesophagus , 1997, Gut.

[130]  Donald Wlodkowic,et al.  Microfluidic single cell arrays to interrogate signalling dynamics of individual, patient-derived hematopoietic stem cells. , 2009, Lab on a chip.

[131]  Kenichi Takahata,et al.  Advances In Micro/Nano Electromechanical Systems And Fabrication Technologies , 2014 .

[132]  N. Teng,et al.  One-step enrichment of nucleated red blood cells. A potential application in perinatal diagnosis. , 1993, Journal of immunological methods.

[133]  Euisik Yoon,et al.  Paired single cell co-culture microenvironments isolated by two-phase flow with continuous nutrient renewal. , 2014, Lab on a chip.

[134]  Thomas J George,et al.  Capture, release and culture of circulating tumor cells from pancreatic cancer patients using an enhanced mixing chip. , 2014, Lab on a chip.

[135]  Ying Li,et al.  CRISPR-Cas9 delivery to hard-to-transfect cells via membrane deformation , 2015, Science Advances.

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

[137]  K. Isselbacher,et al.  Isolation of circulating tumor cells using a microvortex-generating herringbone-chip , 2010, Proceedings of the National Academy of Sciences.

[138]  Luca Berdondini,et al.  Generic technological platform for microfabricating silicon nitride micro- and nanopipette arrays , 2005 .

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

[140]  Jun Wang,et al.  Recent advances in electric analysis of cells in microfluidic systems , 2008, Analytical and bioanalytical chemistry.

[141]  Kidong Park,et al.  Microfluidic multifunctional probe array dielectrophoretic force spectroscopy with wide loading rates. , 2012, ACS nano.

[142]  J Christopher Love,et al.  On-chip activation and subsequent detection of individual antigen-specific T cells. , 2010, Analytical chemistry.

[143]  Thomas Schnelle,et al.  Combined dielectrophoretic field cages and laser tweezers for electrorotation , 2000 .

[144]  James P. Landers,et al.  Toward a Simplified Microfluidic Device for Ultra-Fast Genetic Analysis with Sample-In/Answer-Out Capability: Application to T-Cell Lymphoma Diagnosis , 2008 .

[145]  Fan-Gang Tseng,et al.  Tuning nano electric field to affect restrictive membrane area on localized single cell nano-electroporation , 2013 .

[146]  Robert Langer,et al.  A vector-free microfluidic platform for intracellular delivery , 2013, Proceedings of the National Academy of Sciences.

[147]  S. Leibler,et al.  Bacterial Persistence as a Phenotypic Switch , 2004, Science.

[148]  Melinda J Wilkerson,et al.  Principles and applications of flow cytometry and cell sorting in companion animal medicine. , 2012, The Veterinary clinics of North America. Small animal practice.

[149]  Bernard Lacour,et al.  Circulating trophoblastic cells provide genetic diagnosis in 63 fetuses at risk for cystic fibrosis or spinal muscular atrophy. , 2012, Reproductive biomedicine online.

[150]  Martin Hjort,et al.  Nondestructive nanostraw intracellular sampling for longitudinal cell monitoring , 2017, Proceedings of the National Academy of Sciences.

[151]  Paraskevi Giannakakou,et al.  Functional Characterization of Circulating Tumor Cells with a Prostate-Cancer-Specific Microfluidic Device , 2012, PloS one.

[152]  Dino Di Carlo,et al.  Size-tunable microvortex capture of rare cells. , 2017, Lab on a chip.

[153]  Eugene J. Lim,et al.  Microfluidic, marker-free isolation of circulating tumor cells from blood samples , 2014, Nature Protocols.

[154]  D. Bienzle,et al.  Flow Cytometry in Veterinary Oncology , 2011, Veterinary pathology.

[155]  Yong Huang,et al.  Microfabricated electroporation chip for single cell membrane permeabilization , 2001 .

[156]  T. Graeber,et al.  Circulating tumour cells as a biomarker for diagnosis and staging in pancreatic cancer , 2016, British Journal of Cancer.

[157]  Yuanjin Zhao,et al.  Emerging Droplet Microfluidics. , 2017, Chemical reviews.

[158]  J. Parsons,et al.  CHAPTER 78 – Integrin Signaling: Cell Migration, Proliferation, and Survival , 2003 .

[159]  Richard Heller,et al.  Transfection by Electroporation , 1997, Current protocols in neuroscience.

[160]  Euisik Yoon,et al.  A MICRO/NANO-FLUIDIC CHIP-BASED MICRO-WELL ARRAY FOR HIGH-THROUGHPUT CELL ANALYSIS AND DRUG SCREENING , 2003 .

[161]  Mitch Leslie,et al.  Cell biology. The power of one. , 2011, Science.

[162]  Shuang Hou,et al.  Nanostructure Embedded Microchips for Detection, Isolation, and Characterization of Circulating Tumor Cells , 2014, Accounts of chemical research.

[163]  G. Gradl,et al.  A 3-D microelectrode system for handling and caging single cells and particles , 1999 .

[164]  I-Kao Chiang,et al.  On-chip manipulation of single microparticles, cells, and organisms using surface acoustic waves , 2012, Proceedings of the National Academy of Sciences.

[165]  Jiang.  Optical MEMS for Chemical Analysis and Biomedicine , 2016 .

[166]  Enric Llorens-Bobadilla,et al.  Single-Cell Transcriptomics Reveals a Population of Dormant Neural Stem Cells that Become Activated upon Brain Injury. , 2015, Cell stem cell.

[167]  Fan-Gang Tseng,et al.  Nanolocalized Single-Cell-Membrane Nanoelectroporation: For higher efficiency with high cell viability. , 2014, IEEE Nanotechnology Magazine.

[168]  S. Heimfeld,et al.  Results of retroviral and adenoviral approaches to cancer gene therapy , 1998, Stem cells.

[169]  Arbel Artzy-Schnirman,et al.  Hollow nanoneedle array and its utilization for repeated administration of biomolecules to the same cells. , 2012, ACS nano.

[170]  Leonore A. Herzenberg,et al.  Fetal cells in the blood of pregnant women: detection and enrichment by fluorescence-activated cell sorting. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[171]  Bo Lu,et al.  3D microfilter device for viable circulating tumor cell (CTC) enrichment from blood , 2011, Biomedical microdevices.

[172]  M. Elowitz,et al.  Challenges and emerging directions in single-cell analysis , 2017, Genome Biology.

[173]  Stefan Schinkinger,et al.  Reconfigurable microfluidic integration of a dual-beam laser trap with biomedical applications , 2007, Biomedical microdevices.

[174]  Weiyong Shen,et al.  Laser capture microdissection: from its principle to applications in research on neurodegeneration , 2015, Neural regeneration research.

[175]  N. Mahdieh,et al.  An Overview of Mutation Detection Methods in Genetic Disorders , 2013, Iranian journal of pediatrics.

[176]  Takayuki Shibata,et al.  Development and characterization of hollow microprobe array as a potential tool for versatile and massively parallel manipulation of single cells , 2015, Biomedical Microdevices.

[177]  Luke P. Lee,et al.  A single cell electroporation chip. , 2005, Lab on a chip.

[178]  Salil D. Patel,et al.  Molecular Biotechnology: Therapeutic Applications and Strategies , 1996 .

[179]  M. Rols,et al.  Mechanisms of cell membrane electropermeabilization: a minireview of our present (lack of ?) knowledge. , 2005, Biochimica et biophysica acta.

[180]  Fan-Gang Tseng,et al.  Nanocapillary electrophoretic electrochemical chip: towards analysis of biochemicals released by single cells , 2011, Interface Focus.

[181]  D. Leckband,et al.  Measuring the forces that control protein interactions. , 2000, Annual review of biophysics and biomolecular structure.

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

[183]  Owe Orwar,et al.  Single-cell electroporation , 2010, Analytical and bioanalytical chemistry.

[184]  Kurt Haas,et al.  Single-Cell Electroporationfor Gene Transfer In Vivo , 2001, Neuron.

[185]  Xiaochun Xu,et al.  NanoVelcro Chip for CTC enumeration in prostate cancer patients. , 2013, Methods.

[186]  Takayuki Shibata,et al.  Fabrication of micromanipulator array for cell patterning , 2009 .

[187]  Robert Langer,et al.  Microfluidic squeezing for intracellular antigen loading in polyclonal B-cells as cellular vaccines , 2015, Scientific Reports.

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

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

[190]  Macdara Glynn,et al.  Systems Biology in Single Cells , 2016 .

[191]  Shuang Hou,et al.  Programming Thermoresponsiveness of NanoVelcro Substrates Enables Effective Purification of Circulating Tumor Cells in Lung Cancer Patients , 2014, ACS nano.

[192]  Shuang Hou,et al.  Combined cell surface carbonic anhydrase 9 and CD147 antigens enable high-efficiency capture of circulating tumor cells in clear cell renal cell carcinoma patients , 2016, Oncotarget.

[193]  Jian Wang,et al.  Glioma cell populations grouped by different cell type markers drive brain tumor growth. , 2010, Cancer research.

[194]  David A Hess,et al.  High aldehyde dehydrogenase and expression of cancer stem cell markers selects for breast cancer cells with enhanced malignant and metastatic ability , 2009, Journal of cellular and molecular medicine.

[195]  Michelle D. Wang,et al.  Stretching of single collapsed DNA molecules. , 2000, Biophysical journal.

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

[197]  Grace S. Shieh,et al.  Inferring coregulation of transcription factors and microRNAs in breast cancer. , 2013, Gene.

[198]  Vladimir P. Nikolski,et al.  Transfection of Cardiac Cells by Means of Laser-Assisted Optoporation , 2013 .

[199]  Robert L Sah,et al.  Photo-and Electropatterning of Hydrogel-encapsulated Living Cell Arrays , 2004 .

[200]  Richard P Horgan Mrcog Mrcpi,et al.  ‘Omic’ technologies: genomics, transcriptomics, proteomics and metabolomics , 2011 .

[201]  Hironori Uehara,et al.  mRNA analysis of single living cells , 2003, Journal of nanobiotechnology.

[202]  Y. Ikawa,et al.  A novel method of DNA transfection by laser microbeam cell surgery , 1984 .

[203]  Antony W Burgess,et al.  Epidermal growth factor receptor: mechanisms of activation and signalling. , 2003, Experimental cell research.

[204]  E. Yoon,et al.  High-Throughput Single-Cell Derived Sphere Formation for Cancer Stem-Like Cell Identification and Analysis , 2016, Scientific Reports.

[205]  M. Teitell,et al.  Photothermal nanoblade for large cargo delivery into mammalian cells , 2011, 2011 16th International Solid-State Sensors, Actuators and Microsystems Conference.

[206]  Nozomi Nishimura,et al.  Optoporation and genetic manipulation of cells using femtosecond laser pulses. , 2013, Biophysical journal.

[207]  W. Koh,et al.  Single-cell genome sequencing: current state of the science , 2016, Nature Reviews Genetics.

[208]  Zhen Xu,et al.  Carbon-Nanotube Optoacoustic Lens for Focused Ultrasound Generation and High-Precision Targeted Therapy , 2012, Scientific Reports.

[209]  Shoumei Bai,et al.  Identifying an ovarian cancer cell hierarchy regulated by bone morphogenetic protein 2 , 2015, Proceedings of the National Academy of Sciences.

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

[211]  Majid Minary-Jolandan,et al.  Nanofountain probe electroporation (NFP-E) of single cells. , 2013, Nano letters.

[212]  Daniel Grolimund,et al.  Fast chemical imaging at high spatial resolution by laser ablation inductively coupled plasma mass spectrometry. , 2013, Analytical chemistry.

[213]  Dong Sun,et al.  Single Cell Transfection through Precise Microinjection with Quantitatively Controlled Injection Volumes , 2016, Scientific Reports.

[214]  P. Cullis,et al.  Liposomal drug delivery systems: from concept to clinical applications. , 2013, Advanced drug delivery reviews.

[215]  Shuang Hou,et al.  Clinical Applications of NanoVelcro Rare-Cell Assays for Detection and Characterization of Circulating Tumor Cells , 2016, Theranostics.

[216]  Dong Sun,et al.  Cell manipulation tool with combined microwell array and optical tweezers for cell isolation and deposition , 2013 .

[217]  Amy Brock,et al.  Nanoparticle targeting of anti-cancer drugs that alter intracellular signaling or influence the tumor microenvironment. , 2014, Advanced drug delivery reviews.

[218]  Hyoung Won Baac,et al.  Selective Photomechanical Detachment and Retrieval of Divided Sister Cells from Enclosed Microfluidics for Downstream Analyses. , 2017, ACS nano.

[219]  Fan-Gang Tseng,et al.  Dielectric passivation layer as a substratum on localized single-cell electroporation , 2016 .

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

[221]  Muhammad Waleed,et al.  Single-cell optoporation and transfection using femtosecond laser and optical tweezers. , 2013, Biomedical optics express.

[222]  Unyoung Kim,et al.  Multitarget magnetic activated cell sorter , 2008, Proceedings of the National Academy of Sciences.

[223]  A. Voigt,et al.  Single micro electrode dielectrophoretic tweezers for manipulation of suspended cells and particles. , 1999, Biochimica et biophysica acta.

[224]  Aysun Adan,et al.  Flow cytometry: basic principles and applications , 2017, Critical reviews in biotechnology.

[225]  Zhi Zhu,et al.  Hydrogel Droplet Microfluidics for High-Throughput Single Molecule/Cell Analysis. , 2017, Accounts of chemical research.

[226]  R. Jaenisch,et al.  Microfluidic Control of Cell Pairing and Fusion , 2009, Nature Methods.

[227]  I. Mezić,et al.  Chaotic Mixer for Microchannels , 2002, Science.

[228]  Mahdi Moghimi Zand,et al.  Dielectrophoretic effect of nonuniform electric fields on the protoplast cell , 2017 .

[229]  Shao-Bo Liang,et al.  Application of single-cell technology in cancer research. , 2017, Biotechnology advances.

[230]  Hongbo Xin,et al.  Single Cell Isolation and Analysis , 2016, Front. Cell Dev. Biol..

[231]  Shuang Hou,et al.  Imprinted NanoVelcro Microchips for Isolation and Characterization of Circulating Fetal Trophoblasts: Toward Noninvasive Prenatal Diagnostics. , 2017, ACS nano.

[232]  Yu Sun,et al.  Automated Micropipette Aspiration of Single Cells , 2013, Annals of Biomedical Engineering.

[233]  F. Tang,et al.  Single-cell sequencing in stem cell biology , 2016, Genome Biology.

[234]  Howard Y. Chang,et al.  Transposition of native chromatin for fast and sensitive epigenomic profiling of open chromatin, DNA-binding proteins and nucleosome position , 2013, Nature Methods.

[235]  Fumihito Arai,et al.  High‐speed separation system of randomly suspended single living cells by laser trap and dielectrophoresis , 2001, Electrophoresis.

[236]  Abdur Rahman,et al.  Lab-on-a-chip technology: impacting non-invasive prenatal diagnostics (NIPD) through miniaturisation. , 2014, Lab on a chip.

[237]  David R Walt,et al.  Optical imaging fiber-based single live cell arrays: a high-density cell assay platform. , 2002, Analytical chemistry.

[238]  F. Bragheri,et al.  Integrated microfluidic device for single-cell trapping and spectroscopy , 2013, Scientific Reports.

[239]  Mario R. Capecchi,et al.  High efficiency transformation by direct microinjection of DNA into cultured mammalian cells , 1980, Cell.

[240]  Hong Bin Yang,et al.  Optical detection of single cell lactate release for cancer metabolic analysis. , 2010, Analytical chemistry.

[241]  R. Sandberg,et al.  Single-Cell RNA-Seq Reveals Dynamic, Random Monoallelic Gene Expression in Mammalian Cells , 2014, Science.

[242]  Lakshmi Prasad Dasi,et al.  Force Spectroscopy in the Bloodstream of Live Embryonic Zebrafish with Optical Tweezers , 2014 .

[243]  Xiaolin Wang,et al.  Automated parallel cell isolation and deposition using microwell array and optical tweezers , 2012, 2012 IEEE International Conference on Robotics and Automation.

[244]  R. Tompkins,et al.  Development of a microfabricated cytometry platform for characterization and sorting of individual leukocytes. , 2005, Lab on a chip.

[245]  Gary M. Hieftje,et al.  Nanopipettes: probes for local sample analysis† †Electronic supplementary information (ESI) available. See DOI: 10.1039/c5sc00668f Click here for additional data file. , 2015, Chemical science.

[246]  Hiroyuki Fujita,et al.  Fabrication of Array of Hollow Microcapillaries Used for Injection of Genetic Materials into Animal/Plant Cells , 1999 .

[247]  Steven T. Wereley,et al.  Towards New Methodologies for Manipulation of Colloidal Particles in a Miniaturized Fluidic Device: Optoelectrokinetic Manipulation Technique , 2013 .

[248]  P Neužil,et al.  From chip-in-a-lab to lab-on-a-chip: towards a single handheld electronic system for multiple application-specific lab-on-a-chip (ASLOC). , 2014, Lab on a chip.

[249]  Takao Sugiyama,et al.  Sonoporation: Gene transfer using ultrasound. , 2013, World journal of methodology.

[250]  Frederick R. Cross,et al.  The effects of molecular noise and size control on variability in the budding yeast cell cycle , 2007, Nature.

[251]  Kohji Nakazawa,et al.  Near-IR laser-triggered target cell collection using a carbon nanotube-based cell-cultured substrate. , 2011, ACS nano.

[252]  Tomaso Zambelli,et al.  Isolation of single mammalian cells from adherent cultures by fluidic force microscopy. , 2014, Lab on a chip.

[253]  Li Wang,et al.  Microfluidic device with integrated microfilter of conical-shaped holes for high efficiency and high purity capture of circulating tumor cells , 2014, Scientific Reports.

[254]  Fan-Gang Tseng,et al.  Impact of pulse duration on localized single-cell nano-electroporation. , 2014, The Analyst.

[255]  Marino Zerial,et al.  A decade of molecular cell biology: achievements and challenges , 2011, Nature Reviews Molecular Cell Biology.

[256]  J. Hong,et al.  New tools and new biology: Recent miniaturized systems for molecular and cellular biology , 2013, Molecules and cells.

[257]  A. Murashima,et al.  Gene transduction by sonoporation , 2008, Development, growth & differentiation.

[258]  D. Kent,et al.  High-throughput analysis of single hematopoietic stem cell proliferation in microfluidic cell culture arrays , 2011, Nature Methods.

[259]  A. Felsenfeld.  The power of one. , 2006, Journal of the California Dental Association.

[260]  Christoph A. Merten,et al.  Droplet-based microfluidic platforms for the encapsulation and screening of Mammalian cells and multicellular organisms. , 2008, Chemistry & biology.

[261]  Roberto Osellame,et al.  Hydrodynamic lift for single cell manipulation in a femtosecond laser fabricated optofluidic chip , 2017 .