Dual‐Encoded Microbeads through a Host–Guest Structure: Enormous, Flexible, and Accurate Barcodes for Multiplexed Assays

Bead-based suspension arrays have attracted increasing attention because of the fast growing requirements for high-throughput multiplexed bioassays. Here, novel dual-encoded microbeads (DEBs) based on a host–guest structure are developed. A strawberry-like structure composed of thousands of dye-doped nanoparticles closely packed onto the quantum dot-encoded microbeads is designed and synthesized. Using this 3D structure, the encoded space is fully developed for multicolor encoding. As a result, a 30-barcode library with an outstanding encoding capacity is successfully established and easily decoded via flow cytometry. The separation of organic dyes and quantum dots into two building blocks helps circumvent the optical mutual interference commonly observed in most multicolor barcodes, bringing great convenience for next-step decoding accuracy. As a proof of concept, six typical DEBs are chosen and their feasibility in six-plexed detection for influenza viruses is evaluated. The results show that DEBs can selectively capture the specific targets, with negligible nonspecific absorption; these results thus demonstrate the strong potential of the DEBs as a robust platform for multiplexed bioassays.

[1]  Kang Sun,et al.  Suspension arrays based on nanoparticle-encoded microspheres for high-throughput multiplexed detection. , 2015, Chemical Society reviews.

[2]  Xiaoyuan Chen,et al.  NIR-emitting quantum dot-encoded microbeads through membrane emulsification for multiplexed immunoassays. , 2013, Small.

[3]  Weihong Tan,et al.  Multicolor FRET silica nanoparticles by single wavelength excitation. , 2006, Nano letters.

[4]  C T Lim,et al.  Bead-based microfluidic immunoassays: the next generation. , 2007, Biosensors & bioelectronics.

[5]  Ho Cheung Shum,et al.  Microfluidic generation of multifunctional quantum dot barcode particles. , 2011, Journal of the American Chemical Society.

[6]  W. Chan,et al.  Fabrication of metal nanoshell quantum-dot barcodes for biomolecular detection , 2013 .

[7]  M. Bawendi,et al.  (CdSe)ZnS Core-Shell Quantum Dots - Synthesis and Characterization of a Size Series of Highly Luminescent Nanocrystallites , 1997 .

[8]  Helmuth Möhwald,et al.  Electrostatic Self-Assembly of Silica Nanoparticle−Polyelectrolyte Multilayers on Polystyrene Latex Particles , 1998 .

[9]  I. Willner,et al.  Semiconductor quantum dots for bioanalysis. , 2008, Angewandte Chemie.

[10]  W. Chan,et al.  Engineering multifunctional magnetic-quantum dot barcodes by flow focusing. , 2011, Chemical communications.

[11]  S. Nie,et al.  Quantum-dot-tagged microbeads for multiplexed optical coding of biomolecules , 2001, Nature Biotechnology.

[12]  Baoping Wang,et al.  Multifunctional photonic crystal barcodes from microfluidics , 2012 .

[13]  Jianping Fu,et al.  Encoding through the host-guest structure: construction of multiplexed fluorescent beads. , 2014, Chemical communications.

[14]  Warren C W Chan,et al.  Rapid screening of genetic biomarkers of infectious agents using quantum dot barcodes. , 2011, ACS nano.

[15]  D. Pang,et al.  Fluorescent–magnetic dual-encoded nanospheres: a promising tool for fast-simultaneous-addressable high-throughput analysis , 2012, Nanotechnology.

[16]  Helmuth Möhwald,et al.  Preparation and Characterization of Ordered Nanoparticle and Polymer Composite Multilayers on Colloids , 1999 .

[17]  P. Mulvaney,et al.  The preparation of colloidally stable, water-soluble, biocompatible, semiconductor nanocrystals with a small hydrodynamic diameter. , 2009, ACS nano.

[18]  Alex Rhee,et al.  Facile and rapid one-step mass preparation of quantum-dot barcodes. , 2008, Angewandte Chemie.

[19]  Efficient preparation of magnetic quantum dot barcodes. , 2014, Journal of materials chemistry. B.

[20]  D. Pang,et al.  Optically encoded multifunctional nanospheres for one-pot separation and detection of multiplex DNA sequences. , 2013, Analytical chemistry.

[21]  Xiaohu Gao,et al.  Stable Encapsulation of Quantum Dot Barcodes with Silica Shells , 2010 .

[22]  Y. Chai,et al.  Reverse-micelle synthesis of electrochemically encoded quantum dot barcodes: application to electronic coding of a cancer marker. , 2010, Analytical chemistry.

[23]  G. Sukhorukov,et al.  Toward Encoding Combinatorial Libraries: Charge‐Driven Microencapsulation of Semiconductor Nanocrystals Luminescing in the Visible and Near IR , 2002 .

[24]  M. Bawendi,et al.  Synthesis and characterization of nearly monodisperse CdE (E = sulfur, selenium, tellurium) semiconductor nanocrystallites , 1993 .

[25]  Warren C W Chan,et al.  Automation Highlights from the Literature , 2015, Journal of laboratory automation.

[26]  Kathryn L Kellar,et al.  Multiplexed microsphere-based flow cytometric assays. , 2002, Experimental hematology.

[27]  Jordan J. Feld,et al.  Clinical Validation of Quantum Dot Barcode Diagnostic Technology. , 2016, ACS nano.

[28]  Chaoqing Dong,et al.  Spatially resolved scattering correlation spectroscopy using a total internal reflection configuration. , 2012, Analytical chemistry.

[29]  P. Mulvaney,et al.  The effects of chemisorption on the luminescence of CdSe quantum dots. , 2006, Langmuir : the ACS journal of surfaces and colloids.

[30]  Yong Zhang,et al.  Encapsulation of quantum nanodots in polystyrene and silica micro-/nanoparticles. , 2004, Langmuir : the ACS journal of surfaces and colloids.

[31]  Yong-Kweon Kim,et al.  Surface-enhanced Raman spectroscopic-encoded beads for multiplex immunoassay. , 2007, Journal of combinatorial chemistry.

[32]  Moungi G Bawendi,et al.  In-situ encapsulation of quantum dots into polymer microspheres. , 2006, Langmuir : the ACS journal of surfaces and colloids.

[33]  Yonghong He,et al.  Microfluidic generation of uniform quantum dot-encoded microbeads by gelation of alginate , 2015 .

[34]  Tudor I. Oprea,et al.  Flow cytometry for high-throughput, high-content screening. , 2004, Current opinion in chemical biology.

[35]  Shuming Nie,et al.  Quantum dot-encoded mesoporous beads with high brightness and uniformity: rapid readout using flow cytometry. , 2004, Analytical chemistry.

[36]  David G Spiller,et al.  Encoded microcarriers for high-throughput multiplexed detection. , 2006, Angewandte Chemie.

[37]  Xiebing Wang,et al.  Highly efficient preparation of multiscaled quantum dot barcodes for multiplexed hepatitis B detection. , 2013, ACS nano.

[38]  Z. Gu,et al.  Quantum-dot-coated encoded silica colloidal crystals beads for multiplex coding. , 2009, Chemical communications.

[39]  Walter H. Chang,et al.  Design of an amphiphilic polymer for nanoparticle coating and functionalization. , 2008, Small.

[40]  Xiaoqun Gong,et al.  Self-healing encapsulation strategy for preparing highly stable, functionalized quantum-dot barcodes. , 2014, ACS applied materials & interfaces.

[41]  Huaibin Shen,et al.  Fluorescent QDs-polystyrene composite nanospheres for highly efficient and rapid protein antigen detection , 2013, Journal of Nanoparticle Research.

[42]  J F Keij,et al.  Flow cytometric characterization and classification of multiple dual-color fluorescent microspheres using fluorescence lifetime. , 1998, Cytometry.

[43]  M. Collinson,et al.  Well-Defined Hierarchical Templates for Multimodal Porous Material Fabrication , 2010 .

[44]  R. Nitschke,et al.  Quantum dots versus organic dyes as fluorescent labels , 2008, Nature Methods.

[45]  Igor Nabiev,et al.  Fluorescent nanocrystal-encoded microbeads for multiplexed cancer imaging and diagnosis. , 2008, Critical reviews in oncology/hematology.

[46]  Dieter Stoll,et al.  Protein microarrays for diagnostic assays , 2009, Analytical and bioanalytical chemistry.

[47]  K. Yamaguchi,et al.  Preparation of core–shell composite polymer particles by a novel heterocoagulation based on hydrophobic interaction , 2004 .

[48]  R. G. Freeman,et al.  Submicrometer metallic barcodes. , 2001, Science.

[49]  Homan Kang,et al.  Multilayer fluorescence optically encoded beads for protein detection. , 2010, Analytical biochemistry.

[50]  Mehmet Toner,et al.  Multifunctional Encoded Particles for High-Throughput Biomolecule Analysis , 2007, Science.

[51]  Xiaomei Yan,et al.  Preparation of fluorescence-encoded microspheres in a core–shell structure for suspension arrays , 2010 .

[52]  R J Fulton,et al.  Advanced multiplexed analysis with the FlowMetrix system. , 1997, Clinical chemistry.

[53]  Wang Li,et al.  SERS-fluorescence joint spectral encoding using organic-metal-QD hybrid nanoparticles with a huge encoding capacity for high-throughput biodetection: putting theory into practice. , 2012, Journal of the American Chemical Society.