Multifunctional, inexpensive, and reusable nanoparticle-printed biochip for cell manipulation and diagnosis

Significance Point-of-care diagnostics in the developing world and resource-limited areas require numerous special design considerations to provide effective early detection of disease. Of particular need for these contexts are diagnostic technologies featuring low costs, ease of use, and broad applicability. Here we present a nanoparticle-inkjet-printable microfluidics-based platform that fulfills these criteria and that we expect to significantly reduce the footprint, complexity, and cost of clinical diagnostics. This reusable $0.01 platform is miniaturized to handle small sample volumes and can perform numerous analyses. It can perform complex, minimally invasive analyses of single cells without specialized equipment and personnel. This inexpensive, accessible platform has broad applications in precision diagnostics and is a step toward the democratization of medical technologies. Isolation and characterization of rare cells and molecules from a heterogeneous population is of critical importance in diagnosis of common lethal diseases such as malaria, tuberculosis, HIV, and cancer. For the developing world, point-of-care (POC) diagnostics design must account for limited funds, modest public health infrastructure, and low power availability. To address these challenges, here we integrate microfluidics, electronics, and inkjet printing to build an ultra–low-cost, rapid, and miniaturized lab-on-a-chip (LOC) platform. This platform can perform label-free and rapid single-cell capture, efficient cellular manipulation, rare-cell isolation, selective analytical separation of biological species, sorting, concentration, positioning, enumeration, and characterization. The miniaturized format allows for small sample and reagent volumes. By keeping the electronics separate from microfluidic chips, the former can be reused and device lifetime is extended. Perhaps most notably, the device manufacturing is significantly less expensive, time-consuming, and complex than traditional LOC platforms, requiring only an inkjet printer rather than skilled personnel and clean-room facilities. Production only takes 20 min (vs. up to weeks) and $0.01—an unprecedented cost in clinical diagnostics. The platform works based on intrinsic physical characteristics of biomolecules (e.g., size and polarizability). We demonstrate biomedical applications and verify cell viability in our platform, whose multiplexing and integration of numerous steps and external analyses enhance its application in the clinic, including by nonspecialists. Through its massive cost reduction and usability we anticipate that our platform will enable greater access to diagnostic facilities in developed countries as well as POC diagnostics in resource-poor and developing countries.

[1]  Barbaros Çetin,et al.  Dielectrophoresis in microfluidics technology , 2011, Electrophoresis.

[2]  A. Manz,et al.  Lab-on-a-chip: microfluidics in drug discovery , 2006, Nature Reviews Drug Discovery.

[3]  S. L. Grimes,et al.  The future of clinical engineering: the challenge of change , 2003 .

[4]  J. Voldman Electrical forces for microscale cell manipulation. , 2006, Annual review of biomedical engineering.

[5]  L. Gervais,et al.  Microfluidic Chips for Point‐of‐Care Immunodiagnostics , 2011, Advanced materials.

[6]  H Pertoft,et al.  Fractionation of cells and subcellular particles with Percoll. , 2000, Journal of biochemical and biophysical methods.

[7]  Samuel K Sia,et al.  Lab-on-a-chip devices for global health: past studies and future opportunities. , 2007, Lab on a chip.

[8]  M. Kersaudy-Kerhoas,et al.  Recent advances in microparticle continuous separation. , 2008, IET nanobiotechnology.

[9]  Hongwu Zhu,et al.  Screen-printed microfluidic dielectrophoresis chip for cell separation. , 2015, Biosensors & bioelectronics.

[10]  Jon Richards,et al.  Serial Monitoring of Circulating Tumor Cells Predicts Outcome of Induction Biochemotherapy plus Maintenance Biotherapy for Metastatic Melanoma , 2010, Clinical Cancer Research.

[11]  Andreas Radbruch,et al.  High gradient magnetic cell separation with MACS. , 1990, Cytometry.

[12]  N. Engel,et al.  Point-of-Care Testing for Infectious Diseases: Diversity, Complexity, and Barriers in Low- And Middle-Income Countries , 2012, PLoS medicine.

[13]  Rodrigo Martinez-Duarte,et al.  Microfabrication technologies in dielectrophoresis applications—A review , 2012, Electrophoresis.

[14]  Gianluca Giustolisi,et al.  Theoretical and experimental study of the role of cell-cell dipole interaction in dielectrophoretic devices: application to polynomial electrodes , 2014, Biomedical engineering online.

[15]  K. A. Wolfe,et al.  Microchip-based purification of DNA from biological samples. , 2003, Analytical chemistry.

[16]  Patrick S Daugherty,et al.  Microfluidic protein detection through genetically engineered bacterial cells. , 2006, Journal of proteome research.

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

[18]  Cheng-Hsien Liu,et al.  Dielectrophoresis based‐cell patterning for tissue engineering , 2006, Biotechnology journal.

[19]  Roberto Guerrieri,et al.  A lab-on-a-chip for cell detection and manipulation , 2003 .

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

[21]  R. Sooryakumar,et al.  Manipulation of magnetically labeled and unlabeled cells with mobile magnetic traps. , 2010, Biophysical journal.

[22]  Peter Ndeboc Fonkwo Pricing infectious disease , 2008, EMBO reports.

[23]  H. Morgan,et al.  Ac electrokinetics: a review of forces in microelectrode structures , 1998 .

[24]  Yi-Fang Chen,et al.  Microfluidic chip with microweir structure for continuous sample separating and collecting applications , 2012 .

[25]  Prashanta Dutta,et al.  Dielectrophoretic separation of bioparticles in microdevices: A review , 2014, Electrophoresis.

[26]  Yu Zhang,et al.  Isolation of Circulating Tumor Cells in Patients with Hepatocellular Carcinoma Using a Novel Cell Separation Strategy , 2011, Clinical Cancer Research.

[27]  Mehmet Toner,et al.  A microfluidic device for practical label-free CD4(+) T cell counting of HIV-infected subjects. , 2007, Lab on a chip.

[28]  Ali Khademhosseini,et al.  Nano/Microfluidics for diagnosis of infectious diseases in developing countries. , 2010, Advanced drug delivery reviews.

[29]  S. Gawad,et al.  Impedance spectroscopy flow cytometry: On‐chip label‐free cell differentiation , 2005, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[30]  Helene Andersson,et al.  Microtechnologies and nanotechnologies for single-cell analysis. , 2004, Current opinion in biotechnology.

[31]  Paul Yager,et al.  Cell lysis and protein extraction in a microfluidic device with detection by a fluorogenic enzyme assay. , 2002, Analytical chemistry.

[32]  K. Pienta,et al.  Circulating Tumor Cells Predict Survival Benefit from Treatment in Metastatic Castration-Resistant Prostate Cancer , 2008, Clinical Cancer Research.

[33]  Rashid Bashir,et al.  Dielectrophoresis-based cell manipulation using electrodes on a reusable printed circuit board. , 2009, Lab on a chip.

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

[35]  Richard A Mathies,et al.  Microfluidic devices for DNA sequencing: sample preparation and electrophoretic analysis. , 2003, Current opinion in biotechnology.

[36]  Kristen L. Helton,et al.  Microfluidic Overview of Global Health Issues Microfluidic Diagnostic Technologies for Global Public Health , 2006 .

[37]  Hsueh-Chia Chang,et al.  An integrated dielectrophoretic chip for continuous bioparticle filtering, focusing, sorting, trapping, and detecting. , 2007, Biomicrofluidics.

[38]  J. G. E. Gardeniers,et al.  Lab-On-A-Chip Systems For Biomedical And Environmental Monitoring , 2003, Int. J. Comput. Eng. Sci..

[39]  Alan J Magill,et al.  White blood cell counts and malaria. , 2005, The Journal of infectious diseases.

[40]  Hao Lin,et al.  Low-frequency ac electroporation shows strong frequency dependence and yields comparable transfection results to dc electroporation. , 2012, Journal of controlled release : official journal of the Controlled Release Society.

[41]  Jason P. Gleghorn,et al.  Rare Cell Capture in Microfluidic Devices. , 2011, Chemical engineering science.

[42]  Sam Emaminejad,et al.  Microfluidic diagnostic tool for the developing world: contactless impedance flow cytometry. , 2012, Lab on a chip.

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

[44]  Hansen Bow,et al.  Microfluidics for cell separation , 2010, Medical & Biological Engineering & Computing.

[45]  K. Keddy,et al.  CD4+ lymphocyte count in African patients co-infected with HIV and tuberculosis. , 1995, Journal of acquired immune deficiency syndromes and human retrovirology : official publication of the International Retrovirology Association.

[46]  Curt Balch,et al.  Identification and characterization of ovarian cancer-initiating cells from primary human tumors. , 2008, Cancer research.

[47]  Ali Khademhosseini,et al.  Microfluidics for drug discovery and development: from target selection to product lifecycle management. , 2008, Drug discovery today.

[48]  Zhou Yuan,et al.  Nanotechnology for the detection and kill of circulating tumor cells , 2014, Nanoscale Research Letters.

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

[50]  Charles S Henry,et al.  Advances in microfluidics for environmental analysis. , 2012, The Analyst.

[51]  Fang Yang,et al.  Separation of tumor cells with dielectrophoresis-based microfluidic chip. , 2013, Biomicrofluidics.

[52]  H. O. Fatoyinbo,et al.  Dielectrophoretic separation of Bacillus subtilis spores from environmental diesel particles. , 2007, Journal of environmental monitoring : JEM.

[53]  Jin-Ming Lin,et al.  Particle sorting using a porous membrane in a microfluidic device. , 2011, Lab on a chip.

[54]  Andrew J deMello,et al.  Micro- and nanofluidic systems for high-throughput biological screening. , 2009, Drug discovery today.

[55]  D. Holmes,et al.  Single cell impedance cytometry for identification and counting of CD4 T-cells in human blood using impedance labels. , 2010, Analytical chemistry.

[56]  S. Takayama,et al.  Microfluidics for flow cytometric analysis of cells and particles , 2005, Physiological measurement.

[57]  Burçak Alp,et al.  Building structured biomaterials using AC electrokinetics. , 2003, IEEE engineering in medicine and biology magazine : the quarterly magazine of the Engineering in Medicine & Biology Society.

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

[59]  Michael P. Hughes,et al.  AC electrokinetics: applications for nanotechnology , 2000 .

[60]  Teodor Veres,et al.  Integration and detection of biochemical assays in digital microfluidic LOC devices. , 2010, Lab on a chip.

[61]  Nam-Trung Nguyen,et al.  Rare cell isolation and analysis in microfluidics. , 2014, Lab on a chip.

[62]  Chun-Che Lin,et al.  Sample preconcentration in microfluidic devices , 2011 .

[63]  Kwang Bok Kim,et al.  Red blood cell quantification microfluidic chip using polyelectrolytic gel electrodes , 2009, Electrophoresis.

[64]  Giovanni De Gasperis,et al.  High-frequency electric-field trap for micron and submicron particles , 1995 .