Rapid inertial solution exchange for enrichment and flow cytometric detection of microvesicles.

Exosomes, nanosized membrane-bound vesicles released by cells, play roles in cell signaling, immunology, virology, and oncology. Their study, however, has been hampered by difficulty in isolation and quantification due to their size and the complexity of biological samples. Conventional approaches to improved isolation require specialized equipment and extensive sample preparation time. Therefore, isolation and detection methods of exosomes will benefit biological and clinical studies. Here, we report a microfluidic platform for inline exosome isolation and fluorescent detection using inertial manipulation of antibody-coated exosome capture beads from biological fluids.

[1]  Massimo Spada,et al.  High Levels of Exosomes Expressing CD63 and Caveolin-1 in Plasma of Melanoma Patients , 2009, PloS one.

[2]  Sascha Keller,et al.  Exosomes: from biogenesis and secretion to biological function. , 2006, Immunology letters.

[3]  S. Mathivanan,et al.  Exosomes: extracellular organelles important in intercellular communication. , 2010, Journal of proteomics.

[4]  Cicek Gercel-Taylor,et al.  MicroRNA signatures of tumor-derived exosomes as diagnostic biomarkers of ovarian cancer. , 2008, Gynecologic oncology.

[5]  F. Sánchez‐Madrid,et al.  Intercellular communication: diverse structures for exchange of genetic information , 2012, Nature Reviews Molecular Cell Biology.

[6]  Aram J. Chung,et al.  Three dimensional, sheathless, and high-throughput microparticle inertial focusing through geometry-induced secondary flows. , 2013, Small.

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

[8]  S. Mathivanan,et al.  Two Distinct Populations of Exosomes Are Released from LIM1863 Colon Carcinoma Cell-derived Organoids* , 2012, Molecular & Cellular Proteomics.

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

[10]  Aled Clayton,et al.  Isolation and Characterization of Exosomes from Cell Culture Supernatants and Biological Fluids , 2006, Current protocols in cell biology.

[11]  Mehmet Toner,et al.  Visualization of microscale particle focusing in diluted and whole blood using particle trajectory analysis. , 2012, Lab on a chip.

[12]  H R Büller,et al.  Cell-derived microvesicles and cancer. , 2009, The Netherlands journal of medicine.

[13]  Steven W Graves,et al.  Inertial manipulation and transfer of microparticles across laminar fluid streams. , 2012, Small.

[14]  R. Setterquist,et al.  Exosomes: current knowledge of their composition, biological functions, and diagnostic and therapeutic potentials. , 2012, Biochimica et biophysica acta.

[15]  Gema Moreno-Bueno,et al.  Melanoma exosomes educate bone marrow progenitor cells toward a pro-metastatic phenotype through MET , 2012, Nature Medicine.

[16]  Stefan Sleijfer,et al.  Circulating tumor cells (CTCs): detection methods and their clinical relevance in breast cancer. , 2009, Cancer treatment reviews.

[17]  D. Di Carlo,et al.  Sheathless inertial cell ordering for extreme throughput flow cytometry. , 2010, Lab on a chip.

[18]  H. Shapiro.  Practical Flow Cytometry: Shapiro/Flow Cytometry 4e , 2005 .

[19]  G. Whitesides,et al.  Rapid Prototyping of Microfluidic Systems in Poly(dimethylsiloxane). , 1998, Analytical chemistry.

[20]  Dino Di Carlo,et al.  Microstructure-induced helical vortices allow single-stream and long-term inertial focusing. , 2013, Lab on a chip.

[21]  Graça Raposo,et al.  Exosomal-like vesicles are present in human blood plasma. , 2005, International immunology.

[22]  S. Lee,et al.  Sheathless elasto-inertial particle focusing and continuous separation in a straight rectangular microchannel. , 2011, Lab on a chip.

[23]  James Gimzewski,et al.  Nanostructural and Transcriptomic Analyses of Human Saliva Derived Exosomes , 2010, PloS one.

[24]  Laurence Zitvogel,et al.  Exosomes: composition, biogenesis and function , 2002, Nature Reviews Immunology.

[25]  D. Wong,et al.  Breast Cancer Exosome-like Microvesicles and Salivary Gland Cells Interplay Alters Salivary Gland Cell-Derived Exosome-like Microvesicles In Vitro , 2012, PloS one.

[26]  D. Pisetsky,et al.  The role of microparticles in the pathogenesis of rheumatic diseases , 2010, Nature Reviews Rheumatology.

[27]  Mehmet Toner,et al.  Inertio-elastic focusing of bioparticles in microchannels at high throughput , 2014, Nature Communications.