Counting Single Native Biomolecules and Intact Viruses with Color-Coded Nanoparticles Export

@article{citeulike:2438938, abstract = {Abstract: Nanometer-sized particles such as semiconductor quantum dots and energy-transfer nanoparticles have novel optical properties such as tunable light emission, signal brightness, and multicolor excitation that are not available from traditional organic dyes and fluorescent proteins. Here we report the use of color-coded nanoparticles and dual-color fluorescence coincidence for real-time detection of single native biomolecules and viruses in a microfluidic channel. Using green and red nanoparticles to simultaneously recognize two binding sites on a single target, we demonstrate that individual molecules of genes, proteins, and intact viruses can be detected and identified in complex mixtures without target amplification or probe/target separation. Real-time coincidence analysis of single-photon events allows rapid detection of bound targets and efficient discrimination of excess unbound probes. Quantitative studies indicate that the counting results are remarkably precise when the total numbers of counted molecules are more than 10. The use of bioconjugated nanoparticle probes for single-molecule detection is expected to have important applications in ultrasensitive molecular diagnostics, bioterrorism agent detection, and real-time imaging and tracking of single-molecule processes inside living cells.}, address = {Departments of Biomedical Engineering and Chemistry, Emory University and Georgia Institute of Technology, 101 Woodruff Circle Suite 2001, Atlanta, Georgia 30322, and Centers for Disease Control and Prevention (CDC), and College of Veterinary Medicine, Department of Infectious Diseases, University of Georgia, Athens, Georgia 30602}, author = {Agrawal, A. and Zhang, C. and Byassee, T. and Tripp, R. A. and Nie, S.}, citeulike-article-id = {2438938}, citeulike-linkout-0 = {http://dx.doi.org/10.1021/ac051801t}, citeulike-linkout-1 = {http://pubs.acs.org/doi/abs/10.1021/ac051801t}, day = {15}, doi = {10.1021/ac051801t}, journal = {Anal. Chem.}, keywords = {antibody-nanoparticle-conjugate, ccne-grant, dna-nanoparticle-conjugate, energy-transfer-nanoparticles, fluorescent-quantum-dot, gatech-emory-ccne, oligonucleotide-nanoparticle-conjugate, quantum-dot, single-molecule-sandwich, tnf-alpha}, month = {February}, number = {4}, pages = {1061--1070}, posted-at = {2008-02-27 22:38:11}, priority = {0}, title = {Counting Single Native Biomolecules and Intact Viruses with Color-Coded Nanoparticles}, url = {http://dx.doi.org/10.1021/ac051801t}, volume = {78}, year = {2006} }

TY - JOUR ID - citeulike:2438938 L3 - citeulike-article-id:2438938 N2 - Abstract: Nanometer-sized particles such as semiconductor quantum dots and energy-transfer nanoparticles have novel optical properties such as tunable light emission, signal brightness, and multicolor excitation that are not available from traditional organic dyes and fluorescent proteins. Here we report the use of color-coded nanoparticles and dual-color fluorescence coincidence for real-time detection of single native biomolecules and viruses in a microfluidic channel. Using green and red nanoparticles to simultaneously recognize two binding sites on a single target, we demonstrate that individual molecules of genes, proteins, and intact viruses can be detected and identified in complex mixtures without target amplification or probe/target separation. Real-time coincidence analysis of single-photon events allows rapid detection of bound targets and efficient discrimination of excess unbound probes. Quantitative studies indicate that the counting results are remarkably precise when the total numbers of counted molecules are more than 10. The use of bioconjugated nanoparticle probes for single-molecule detection is expected to have important applications in ultrasensitive molecular diagnostics, bioterrorism agent detection, and real-time imaging and tracking of single-molecule processes inside living cells. IS - 4 JF - Anal. Chem. AD - Departments of Biomedical Engineering and Chemistry, Emory University and Georgia Institute of Technology, 101 Woodruff Circle Suite 2001, Atlanta, Georgia 30322, and Centers for Disease Control and Prevention (CDC), and College of Veterinary Medicine, Department of Infectious Diseases, University of Georgia, Athens, Georgia 30602 EP - 1070 TI - Counting Single Native Biomolecules and Intact Viruses with Color-Coded Nanoparticles VL - 78 SP - 1061 KW - antibody-nanoparticle-conjugate KW - ccne-grant KW - dna-nanoparticle-conjugate KW - energy-transfer-nanoparticles KW - fluorescent-quantum-dot KW - gatech-emory-ccne KW - oligonucleotide-nanoparticle-conjugate KW - quantum-dot KW - single-molecule-sandwich KW - tnf-alpha AU - Agrawal, A AU - Zhang, C AU - Byassee, T AU - Tripp, RA AU - Nie, S PY - 2006/02/15/ UR - http://dx.doi.org/10.1021/ac051801t ER -