Microfluidic Screening of Circulating Tumor Biomarkers toward Liquid Biopsy

The development of early and personalized diagnostic protocol with rapid response and high accuracy is considered the most promising avenue to advance point-of-care testing for tumor diagnosis and therapy. Given the growing awareness of the limitations of conventional tissue biopsy for gathering tumor information, considerable interest has recently been aroused in liquid biopsy. Among a myriad of analytical approaches proposed for liquid biopsy, microfluidics-based separation and purification techniques possess merits of high throughput, low samples consumption, high flexibility, low cost, high sensitivity, automation capability and enhanced spatio-temporal control. These characteristics endow microfluidics to serve as an emerging and promising tool in tumor diagnosis and prognosis by identifying specific circulating tumor biomarkers. In this review, we will put our focus on three key categories of circulating tumor biomarkers, namely, circulating tumor cells (CTCs), circulating exosomes, and circulating nucleic acids (cNAs), and discuss the significant roles of microfluidics in the separation and analysis of circulating tumor biomarkers. Recent advances in microfluidic separation and analysis of CTCs, exosomes, and cNAs will be highlighted and tabulated. Finally, the current challenges and future niches of using microfluidic techniques in the separation and analysis of circulating tumor biomarkers will be discussed.

[1]  C. Puleo,et al.  Microfluidic means of achieving attomolar detection limits with molecular beacon probes. , 2009, Lab on a chip.

[2]  Hongtao Feng,et al.  High throughput capture of circulating tumor cells using an integrated microfluidic system. , 2013, Biosensors & bioelectronics.

[3]  Susana Cardoso,et al.  Implementing a strategy for on-chip detection of cell-free DNA fragments using GMR sensors: A translational application in cancer diagnostics using ALU elements , 2016 .

[4]  F F Becker,et al.  Cell separation on microfabricated electrodes using dielectrophoretic/gravitational field-flow fractionation. , 1999, Analytical chemistry.

[5]  Linlin Li,et al.  Roles of particle size, shape and surface chemistry of mesoporous silica nanomaterials on biological systems , 2017 .

[6]  T. Huang,et al.  Acoustic separation of circulating tumor cells , 2015, Proceedings of the National Academy of Sciences.

[7]  H. Amini,et al.  Label-free cell separation and sorting in microfluidic systems , 2010, Analytical and bioanalytical chemistry.

[8]  S. Kalams,et al.  DC-Dielectrophoretic separation of biological cells by size , 2008, Biomedical microdevices.

[9]  Jeong-Gun Lee,et al.  SSA-MOA: a novel CTC isolation platform using selective size amplification (SSA) and a multi-obstacle architecture (MOA) filter. , 2012, Lab on a chip.

[10]  Dino Di Carlo,et al.  High-throughput size-based rare cell enrichment using microscale vortices. , 2011, Biomicrofluidics.

[11]  Minseok S Kim,et al.  A trachea-inspired bifurcated microfilter capturing viable circulating tumor cells via altered biophysical properties as measured by atomic force microscopy. , 2013, Small.

[12]  Byungkyu Kim,et al.  Separation of malignant human breast cancer epithelial cells from healthy epithelial cells using an advanced dielectrophoresis-activated cell sorter (DACS) , 2009, Analytical and bioanalytical chemistry.

[13]  James N Turner,et al.  Isolation of tumor cells using size and deformation. , 2009, Journal of chromatography. A.

[14]  Fang Yang,et al.  Dielectrophoretic separation of colorectal cancer cells. , 2010, Biomicrofluidics.

[15]  H. Jung,et al.  Continuous labeling of circulating tumor cells with microbeads using a vortex micromixer for highly selective isolation. , 2013, Biosensors & bioelectronics.

[16]  김승일.  A Trachea-Inspired Bifurcated Microfilter Capturing Viable Circulating Tumor Cells via Altered Biophysical Properties as Measured by Atomic Force Microscopy , 2013 .

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

[18]  F. Becker,et al.  Isolation of rare cells from cell mixtures by dielectrophoresis , 2009, Electrophoresis.

[19]  Jaesung Park,et al.  Microfluidic fabrication of cell-derived nanovesicles as endogenous RNA carriers. , 2014, Lab on a chip.

[20]  Jina Ko,et al.  Detection and isolation of circulating exosomes and microvesicles for cancer monitoring and diagnostics using micro-/nano-based devices. , 2016, The Analyst.

[21]  Sonal Patel,et al.  A single-molecule method for the quantitation of microRNA gene expression , 2005, Nature Methods.

[22]  Y. Lo,et al.  A Single-Cell Assay for Time Lapse Studies of Exosome Secretion and Cell Behaviors. , 2016, Small.

[23]  Akira Ono,et al.  Size-Based Isolation of Circulating Tumor Cells in Lung Cancer Patients Using a Microcavity Array System , 2013, PloS one.

[24]  Minseok S Kim,et al.  Highly efficient assay of circulating tumor cells by selective sedimentation with a density gradient medium and microfiltration from whole blood. , 2012, Analytical chemistry.

[25]  Chung-Liang Ho,et al.  Noninvasive saliva-based EGFR gene mutation detection in patients with lung cancer. , 2014, American journal of respiratory and critical care medicine.

[26]  John X. J. Zhang,et al.  Microscale Magnetic Field Modulation for Enhanced Capture and Distribution of Rare Circulating Tumor Cells , 2015, Scientific Reports.

[27]  Kazunori Hoshino,et al.  Multiscale immunomagnetic enrichment of circulating tumor cells: from tubes to microchips. , 2014, Lab on a chip.

[28]  Yang Yang,et al.  A microfluidic ExoSearch chip for multiplexed exosome detection towards blood-based ovarian cancer diagnosis. , 2016, Lab on a chip.

[29]  Byungkyu Kim,et al.  An efficient cell separation system using 3D-asymmetric microelectrodes. , 2005, Lab on a chip.

[30]  Ion Stiharu,et al.  Interdigitated comb‐like electrodes for continuous separation of malignant cells from blood using dielectrophoresis , 2011, Electrophoresis.

[31]  Hakho Lee,et al.  Acoustic purification of extracellular microvesicles. , 2015, ACS nano.

[32]  Daniel B. Martin,et al.  Circulating microRNAs as stable blood-based markers for cancer detection , 2008, Proceedings of the National Academy of Sciences.

[33]  Zhiyuan Hu,et al.  Label-Free Quantitative Detection of Tumor-Derived Exosomes through Surface Plasmon Resonance Imaging , 2014, Analytical chemistry.

[34]  Tomoko Yoshino,et al.  Size-selective microcavity array for rapid and efficient detection of circulating tumor cells. , 2010, Analytical chemistry.

[35]  James R Heath,et al.  Microfluidics-based single-cell functional proteomics for fundamental and applied biomedical applications. , 2014, Annual review of analytical chemistry.

[36]  Kelvin J. Liu,et al.  A surface topography assisted droplet manipulation platform for biomarker detection and pathogen identification. , 2011, Lab on a chip.

[37]  Gwo-Bin Lee,et al.  Automatic microfluidic platform for cell separation and nucleus collection , 2007, Biomedical microdevices.

[38]  R. Pethig,et al.  ApoStream(™), a new dielectrophoretic device for antibody independent isolation and recovery of viable cancer cells from blood. , 2012, Biomicrofluidics.

[39]  Peng Chen,et al.  Computational analysis of microfluidic immunomagnetic rare cell separation from a particulate blood flow. , 2012, Analytical chemistry.

[40]  Carla Oliveira,et al.  Evidence-Based Clinical Use of Nanoscale Extracellular Vesicles in Nanomedicine. , 2016, ACS nano.

[41]  Siyang Zheng,et al.  Membrane microfilter device for selective capture, electrolysis and genomic analysis of human circulating tumor cells. , 2007, Journal of chromatography. A.

[42]  Nobuyuki Yamamoto,et al.  Microcavity array system for size-based enrichment of circulating tumor cells from the blood of patients with small-cell lung cancer. , 2013, Analytical chemistry.

[43]  Ciprian Iliescu,et al.  Label-free isolation of circulating tumor cells in microfluidic devices: Current research and perspectives. , 2013, Biomicrofluidics.

[44]  Donald E Ingber,et al.  A combined micromagnetic-microfluidic device for rapid capture and culture of rare circulating tumor cells. , 2012, Lab on a chip.

[45]  Hakho Lee,et al.  Protein typing of circulating microvesicles allows real-time monitoring of glioblastoma therapy , 2012, Nature Medicine.

[46]  M. Stroun,et al.  Neoplastic characteristics of the DNA found in the plasma of cancer patients. , 1989, Oncology.

[47]  Suhong Yu,et al.  Isolation and characterization of living circulating tumor cells in patients by immunomagnetic negative enrichment coupled with flow cytometry , 2015, Cancer.

[48]  K. Pantel,et al.  Challenges in circulating tumour cell research , 2014, Nature Reviews Cancer.

[49]  D. Go,et al.  On-chip surface acoustic wave lysis and ion-exchange nanomembrane detection of exosomal RNA for pancreatic cancer study and diagnosis. , 2015, Lab on a chip.

[50]  Chang-Yu Chen,et al.  Separation and detection of rare cells in a microfluidic disk via negative selection. , 2011, Lab on a chip.

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

[52]  Michael J Heller,et al.  Low level epifluorescent detection of nanoparticles and DNA on dielectrophoretic microarrays , 2014, Journal of biophotonics.

[53]  R. Cerione,et al.  Microfluidic isolation of cancer-cell-derived microvesicles from hetergeneous extracellular shed vesicle populations , 2014, Biomedical Microdevices.

[54]  Joseph D'Silva,et al.  Deterministic separation of cancer cells from blood at 10 mL/min. , 2012, AIP advances.

[55]  Maciej Zborowski,et al.  Rare cell separation and analysis by magnetic sorting. , 2011, Analytical chemistry.

[56]  A. Bardelli,et al.  Blood circulating tumor DNA for non‐invasive genotyping of colon cancer patients , 2016, Molecular oncology.

[57]  Paolo Bergese,et al.  Colorimetric nanoplasmonic assay to determine purity and titrate extracellular vesicles. , 2015, Analytical chemistry.

[58]  Daniel T Chiu,et al.  Deformability considerations in filtration of biological cells. , 2010, Lab on a chip.

[59]  Xiaojing Zhang,et al.  Microsystems for controlled genetic perturbation of live Drosophila embryos: RNA interference, development robustness and drug screening , 2009 .

[60]  T. Wurdinger,et al.  Microfluidic isolation and transcriptome analysis of serum microvesicles. , 2010, Lab on a chip.

[61]  P. Kwok,et al.  Genome mapping on nanochannel arrays for structural variation analysis and sequence assembly , 2012, Nature Biotechnology.

[62]  Francis Barany,et al.  High-throughput selection, enumeration, electrokinetic manipulation, and molecular profiling of low-abundance circulating tumor cells using a microfluidic system. , 2011, Analytical chemistry.

[63]  H. Jung,et al.  Continuous separation of breast cancer cells from blood samples using multi-orifice flow fractionation (MOFF) and dielectrophoresis (DEP). , 2011, Lab on a chip.

[64]  Han Wei Hou,et al.  Pinched flow coupled shear-modulated inertial microfluidics for high-throughput rare blood cell separation. , 2011, Lab on a chip.

[65]  Massimo Cristofanilli,et al.  Dielectric cell separation of fine needle aspirates from tumor xenografts. , 2008, Journal of separation science.

[66]  Fei Huang,et al.  Rapid isolation of cancer cells using microfluidic deterministic lateral displacement structure. , 2013, Biomicrofluidics.

[67]  James P Landers,et al.  Disposable polyester-toner electrophoresis microchips for DNA analysis. , 2012, The Analyst.

[68]  Peng Chen,et al.  Inkjet-Print Micromagnet Array on Glass Slides for Immunomagnetic Enrichment of Circulating Tumor Cells , 2016, Annals of Biomedical Engineering.

[69]  Kyall J. Pocock,et al.  Efficient microfluidic negative enrichment of circulating tumor cells in blood using roughened PDMS. , 2015, The Analyst.

[70]  J. Dear,et al.  Quantification of human urinary exosomes by nanoparticle tracking analysis , 2014 .

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

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

[73]  A. Lee,et al.  Droplet microfluidics for amplification-free genetic detection of single cells. , 2012, Lab on a chip.

[74]  K. Chi.  The tumour trail left in blood , 2016, Nature.

[75]  G. Spoto,et al.  Biosensors for liquid biopsy: circulating nucleic acids to diagnose and treat cancer , 2016, Analytical and Bioanalytical Chemistry.

[76]  Klaus Pantel,et al.  Tumor metastasis: moving new biological insights into the clinic , 2013, Nature Medicine.

[77]  H. Lilja,et al.  Microfluidic, label-free enrichment of prostate cancer cells in blood based on acoustophoresis. , 2012, Analytical chemistry.

[78]  Y. Huang,et al.  Cell separation by dielectrophoretic field-flow-fractionation. , 2000, Analytical chemistry.

[79]  Mark Burns,et al.  Microfluidic chemical analysis systems. , 2011, Annual review of chemical and biomolecular engineering.

[80]  Ron R Lin,et al.  High-throughput single-molecule optofluidic analysis , 2011, Nature Methods.

[81]  Weihong Tan,et al.  Enrichment of cancer cells using aptamers immobilized on a microfluidic channel. , 2009, Analytical chemistry.

[82]  Kyung-A Hyun,et al.  Isolation and enrichment of circulating biomarkers for cancer screening, detection, and diagnostics. , 2016, The Analyst.

[83]  Yong Zeng,et al.  Microfluidic Exosome Analysis toward Liquid Biopsy for Cancer , 2016, Journal of laboratory automation.

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

[85]  Ronald Pethig,et al.  The removal of human leukaemia cells from blood using interdigitated microelectrodes , 1994 .

[86]  Antje J Baeumner,et al.  Microfluidic isolation of nucleic acids. , 2014, Angewandte Chemie.

[87]  P. Lisowski,et al.  Microfluidic Paper-Based Analytical Devices (μPADs) and Micro Total Analysis Systems (μTAS): Development, Applications and Future Trends , 2013, Chromatographia.

[88]  Daniel T Chiu,et al.  Sensitive and high-throughput isolation of rare cells from peripheral blood with ensemble-decision aliquot ranking. , 2012, Angewandte Chemie.

[89]  M. Vellekoop,et al.  Guided Dielectrophoresis: A Robust Method for Continuous Particle and Cell Separation , 2010, IEEE Sensors Journal.

[90]  Mehmet Toner,et al.  Antibody-functionalized fluid-permeable surfaces for rolling cell capture at high flow rates. , 2012, Biophysical journal.

[91]  Brian N. Johnson,et al.  An integrated nanoliter DNA analysis device. , 1998, Science.

[92]  Peter R C Gascoyne,et al.  Dielectrophoretic Separation of Cancer Cells from Blood. , 1997, IEEE transactions on industry applications.

[93]  Sudhir Srivastava,et al.  A simple packed bed device for antibody labelled rare cell capture from whole blood. , 2012, Lab on a chip.

[94]  C. Lim,et al.  Isolation and retrieval of circulating tumor cells using centrifugal forces , 2013, Scientific Reports.

[95]  Kazunori Hoshino,et al.  Microchip-based immunomagnetic detection of circulating tumor cells. , 2011, Lab on a chip.

[96]  Peng Chen,et al.  Versatile immunomagnetic nanocarrier platform for capturing cancer cells. , 2013, ACS nano.

[97]  Swee Jin Tan,et al.  Microdevice for the isolation and enumeration of cancer cells from blood , 2009, Biomedical microdevices.

[98]  Fangqiong Tang,et al.  Shape matters when engineering mesoporous silica-based nanomedicines. , 2016, Biomaterials science.

[99]  B. Greene,et al.  Microtube device for selectin-mediated capture of viable circulating tumor cells from blood. , 2012, Clinical chemistry.

[100]  P Mandel,et al.  Les acides nucleiques du plasma sanguin chez l' homme , 1948 .

[101]  Akira Matsumoto,et al.  A label-free electrical detection of exosomal microRNAs using microelectrode array. , 2012, Chemical communications.

[102]  Jaesung Park,et al.  Microfluidic filtration system to isolate extracellular vesicles from blood. , 2012, Lab on a chip.

[103]  C. Lim,et al.  DEAN FLOW FRACTIONATION (DFF) ISOLATION OF CIRCULATING TUMOR CELLS (CTCs) FROM BLOOD , 2011 .

[104]  Chaoyong James Yang,et al.  Massively parallel single-molecule and single-cell emulsion reverse transcription polymerase chain reaction using agarose droplet microfluidics. , 2012, Analytical chemistry.

[105]  Kazuo Takeda,et al.  Enumeration, characterization, and collection of intact circulating tumor cells by cross contamination‐free flow cytometry , 2011, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[106]  A. Heeger,et al.  Selection of mammalian cells based on their cell-cycle phase using dielectrophoresis , 2007, Proceedings of the National Academy of Sciences of the United States of America.

[107]  Peter R C Gascoyne,et al.  Dielectrophoretic segregation of different human cell types on microscope slides. , 2005, Analytical chemistry.

[108]  Kazunori Hoshino,et al.  Immunomagnetic nanoscreening of circulating tumor cells with a motion controlled microfluidic system , 2012, Biomedical Microdevices.

[109]  Eva M. Schmelz,et al.  Selective concentration of human cancer cells using contactless dielectrophoresis , 2011, Electrophoresis.

[110]  T. Huang,et al.  Cell separation using tilted-angle standing surface acoustic waves , 2014, Proceedings of the National Academy of Sciences.

[111]  F F Becker,et al.  The removal of human breast cancer cells from hematopoietic CD34+ stem cells by dielectrophoretic field-flow-fractionation. , 1999, Journal of Hematotherapy & Stem Cell Research.

[112]  Peter C. Y. Chen,et al.  Slanted spiral microfluidics for the ultra-fast, label-free isolation of circulating tumor cells. , 2014, Lab on a chip.

[113]  Hong Wu,et al.  Three-dimensional nanostructured substrates toward efficient capture of circulating tumor cells. , 2009, Angewandte Chemie.

[114]  L. Mazutis,et al.  Quantitative and sensitive detection of rare mutations using droplet-based microfluidics. , 2011, Lab on a chip.

[115]  Rajan P Kulkarni,et al.  Rapid inertial solution exchange for enrichment and flow cytometric detection of microvesicles. , 2015, Biomicrofluidics.

[116]  Xingyu Jiang,et al.  Size-based hydrodynamic rare tumor cell separation in curved microfluidic channels. , 2013, Biomicrofluidics.

[117]  A. B. Frazier,et al.  Microsystems for isolation and electrophysiological analysis of breast cancer cells from blood. , 2006, Biosensors & bioelectronics.

[118]  D. Poenar,et al.  An integrated on-chip platform for negative enrichment of tumour cells. , 2016, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[119]  Diane M Simeone,et al.  Microfluidic device (ExoChip) for on-chip isolation, quantification and characterization of circulating exosomes. , 2014, Lab on a chip.

[120]  Martin Fuchs,et al.  DNA mapping using microfluidic stretching and single-molecule detection of fluorescent site-specific tags. , 2004, Genome research.

[121]  Andrea Toma,et al.  Breaking the diffusion limit with super-hydrophobic delivery of molecules to plasmonic nanofocusing SERS structures , 2011 .

[122]  Tza-Huei Wang,et al.  Analysis of single nucleic acid molecules in micro- and nano-fluidics. , 2016, Lab on a chip.

[123]  Saeid Nahavandi,et al.  Microfluidic platforms for biomarker analysis. , 2014, Lab on a chip.

[124]  Peng Zhang,et al.  Ultrasensitive microfluidic analysis of circulating exosomes using a nanostructured graphene oxide/polydopamine coating. , 2016, Lab on a chip.

[125]  D. Di Carlo.  Inertial microfluidics. , 2009, Lab on a chip.

[126]  Weihong Tan,et al.  Aptamer-based microfluidic device for enrichment, sorting, and detection of multiple cancer cells. , 2009, Analytical chemistry.

[127]  Peng Chen,et al.  Screening and Molecular Analysis of Single Circulating Tumor Cells Using Micromagnet Array , 2015, Scientific Reports.

[128]  Tza-Huei Wang,et al.  Decoding circulating nucleic acids in human serum using microfluidic single molecule spectroscopy. , 2010, Journal of the American Chemical Society.

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

[130]  Gwo-Bin Lee,et al.  High-purity and label-free isolation of circulating tumor cells (CTCs) in a microfluidic platform by using optically-induced-dielectrophoretic (ODEP) force. , 2013, Lab on a chip.

[131]  Chiun-Sheng Huang,et al.  Enumeration and viability of rare cells in a microfluidic disk via positive selection approach. , 2012, Analytical biochemistry.

[132]  S. Muller,et al.  Polymer-monovalent salt-induced DNA compaction studied via single-molecule microfluidic trapping. , 2012, Lab on a chip.

[133]  Matt Trau,et al.  Detecting exosomes specifically: a multiplexed device based on alternating current electrohydrodynamic induced nanoshearing. , 2014, Analytical chemistry.

[134]  Hakho Lee,et al.  Label-free detection and molecular profiling of exosomes with a nano-plasmonic sensor , 2014, Nature Biotechnology.

[135]  O. Warburg.  [Origin of cancer cells]. , 1956, Oncologia.

[136]  F F Becker,et al.  Separation of human breast cancer cells from blood by differential dielectric affinity. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[137]  Antje J. Baeumner,et al.  Design and fabrication of a microfluidic device for near-single cell mRNA isolation using a copper hot embossing master , 2009 .

[138]  M. Sano,et al.  Contactless dielectrophoresis: a new technique for cell manipulation , 2009, Biomedical microdevices.

[139]  Mehmet Toner,et al.  Inertial Focusing for Tumor Antigen–Dependent and –Independent Sorting of Rare Circulating Tumor Cells , 2013, Science Translational Medicine.

[140]  Chao Liu,et al.  Double spiral microchannel for label-free tumor cell separation and enrichment. , 2012, Lab on a chip.

[141]  Cesar M. Castro,et al.  Magnetic nanosensor for detection and profiling of erythrocyte-derived microvesicles. , 2013, ACS nano.

[142]  Chwee Teck Lim,et al.  Versatile label free biochip for the detection of circulating tumor cells from peripheral blood in cancer patients. , 2010, Biosensors & bioelectronics.

[143]  V. Ambros,et al.  The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14 , 1993, Cell.

[144]  Martin A. M. Gijs,et al.  Selective breast cancer cell capture, culture, and immunocytochemical analysis using self-assembled magnetic bead patterns in a microfluidic chip. , 2010, Langmuir : the ACS journal of surfaces and colloids.

[145]  Graça Minas,et al.  Biomedical microfluidic devices by using low-cost fabrication techniques: A review. , 2016, Journal of biomechanics.

[146]  Juan J de Pablo,et al.  A microfluidic system for large DNA molecule arrays. , 2004, Analytical chemistry.

[147]  M. Gijs,et al.  Droplet-based DNA purification in a magnetic lab-on-a-chip. , 2006, Angewandte Chemie.

[148]  John X. J. Zhang,et al.  Magnetic‐Nanoparticle‐Based Immunoassays‐on‐Chip: Materials Synthesis, Surface Functionalization, and Cancer Cell Screening , 2016 .

[149]  G Medoro,et al.  Levitation and movement of human tumor cells using a printed circuit board device based on software-controlled dielectrophoresis. , 2003, Biotechnology and bioengineering.

[150]  Byungkyu Kim,et al.  Label-free, microfluidic separation and enrichment of human breast cancer cells by adhesion difference. , 2007, Lab on a chip.

[151]  S. Soper,et al.  A titer plate-based polymer microfluidic platform for high throughput nucleic acid purification , 2008, Biomedical microdevices.

[152]  M. Heller,et al.  Isolation of cultured cervical carcinoma cells mixed with peripheral blood cells on a bioelectronic chip. , 1998, Analytical chemistry.

[153]  Poenar Daniel Puiu,et al.  Microfluidic platform for negative enrichment of circulating tumor cells , 2014, Biomedical Microdevices.

[154]  A. Baeumner,et al.  Isolation and amplification of mRNA within a simple microfluidic lab on a chip. , 2014, Analytical chemistry.

[155]  Dino Di Carlo,et al.  Automated cellular sample preparation using a Centrifuge-on-a-Chip. , 2011, Lab on a chip.

[156]  M. Akpinar-Elci,et al.  Ac ce pte d M an us cri pt 1 1 Upper Airways diseases in Agriculture United Airways Disease Among Crop Farmers , 2016 .

[157]  Ali Beskok,et al.  Dielectrophoretic separation of mouse melanoma clones. , 2010, Biomicrofluidics.

[158]  Inhak Choi,et al.  Circulating tumor cell microseparator based on lateral magnetophoresis and immunomagnetic nanobeads. , 2013, Analytical chemistry.

[159]  Bert Vogelstein,et al.  DETECTION OF CIRCULATING TUMOR DNA IN EARLY AND LATE STAGE HUMAN MALIGNANCIES , 2014 .

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

[161]  Richard O Hynes,et al.  Metastatic cells will take any help they can get. , 2011, Cancer cell.

[162]  R. Pethig.  Review article-dielectrophoresis: status of the theory, technology, and applications. , 2010, Biomicrofluidics.

[163]  Lidong Qin,et al.  A robotics platform for automated batch fabrication of high density, microfluidics-based DNA microarrays, with applications to single cell, multiplex assays of secreted proteins. , 2011, The Review of scientific instruments.

[164]  H. Duan,et al.  From structures to functions: insights into exosomes as promising drug delivery vehicles. , 2016, Biomaterials science.

[165]  Evelyn Wenkel,et al.  Circulating Micro-RNAs as Potential Blood-Based Markers for Early Stage Breast Cancer Detection , 2012, PloS one.

[166]  Biana Godin,et al.  Ciliated micropillars for the microfluidic-based isolation of nanoscale lipid vesicles. , 2013, Lab on a chip.

[167]  S. Kang,et al.  Microchip‐Based Capillary Electrophoresis for DNA Analysis in Modern Biotechnology: A Review , 2009 .

[168]  Juan J de Pablo,et al.  Elongation and migration of single DNA molecules in microchannels using oscillatory shear flows. , 2009, Lab on a chip.

[169]  J H Myung,et al.  Microfluidic devices to enrich and isolate circulating tumor cells. , 2015, Lab on a chip.

[170]  Bob S. Carter,et al.  Chip-based analysis of exosomal mRNA mediating drug resistance in glioblastoma , 2015, Nature Communications.

[171]  Michael J Heller,et al.  Dielectrophoretic isolation and detection of cfc‐DNA nanoparticulate biomarkers and virus from blood , 2013, Electrophoresis.

[172]  Salvatore Campo,et al.  Circulating microRNAs: new biomarkers in diagnosis, prognosis and treatment of cancer (review). , 2012, International journal of oncology.

[173]  Clotilde Théry,et al.  Biogenesis and secretion of exosomes. , 2014, Current opinion in cell biology.

[174]  Mingzhou Guo,et al.  Spatially gradated segregation and recovery of circulating tumor cells from peripheral blood of cancer patients. , 2013, Biomicrofluidics.

[175]  Kyung-A Hyun,et al.  Negative enrichment of circulating tumor cells using a geometrically activated surface interaction chip. , 2013, Analytical chemistry.

[176]  M. Heller,et al.  Dielectrophoretic cell separation and gene expression profiling on microelectronic chip arrays. , 2002, Analytical chemistry.

[177]  Kazunori Hoshino,et al.  Microfluidic immunodetection of cancer cells via site-specific microcontact printing of antibodies on nanoporous surface. , 2013, Methods.

[178]  Kyung-A Hyun,et al.  Continual collection and re-separation of circulating tumor cells from blood using multi-stage multi-orifice flow fractionation. , 2013, Biomicrofluidics.

[179]  Ulrich Keilholz,et al.  Negative enrichment by immunomagnetic nanobeads for unbiased characterization of circulating tumor cells from peripheral blood of cancer patients , 2010, Journal of Translational Medicine.

[180]  Paul I. Okagbare,et al.  Highly efficient circulating tumor cell isolation from whole blood and label-free enumeration using polymer-based microfluidics with an integrated conductivity sensor. , 2008, Journal of the American Chemical Society.

[181]  Y. Huang,et al.  Introducing dielectrophoresis as a new force field for field-flow fractionation. , 1997, Biophysical journal.