Superlocalization of single molecules and nanoparticles in high-fidelity optical imaging microfluidic devices.

Superlocalization of single molecules and nanoparticles with a precision of subnanometer to a few tens of nanometers is crucial for elucidating nanoscale structures and movements in biological and chemical systems. A novel design of ultraflat and ultrathin glass/polydimethylsiloxane (PDMS) hybrid microdevices is introduced to provide almost uncompromised optical imaging quality for on-chip superlocalization and super-resolution imaging of single molecules and nanoparticles under a variety of microscopy modes. The performance of the high-fidelity (Hi-Fi) optical imaging microfluidic device was validated by precisely mapping micronecklaces made of fluorescent microtubules and 40 nm gold nanoparticles and by demonstrating the activation and excitation cycles of single Alexa Fluor 647 dyes for direct stochastic optical reconstruction microscopy in PDMS-based microchannels for the first time. Furthermore, the microdevice's feasibility for multimodality microscopy imaging was demonstrated by a vertical scan of live cells in epi-fluorescence and differential interference contrast (DIC) microscopy modes simultaneously.

[1]  K. Jensen,et al.  Cells on chips , 2006, Nature.

[2]  P. Schultz,et al.  Microfluidic device for single-molecule experiments with enhanced photostability. , 2009, Journal of the American Chemical Society.

[3]  Shu Chien,et al.  Fluorescence proteins, live-cell imaging, and mechanobiology: seeing is believing. , 2008, Annual review of biomedical engineering.

[4]  Mark Bates,et al.  Super-resolution fluorescence microscopy. , 2009, Annual review of biochemistry.

[5]  Paul J. Choi,et al.  Quantifying E. coli Proteome and Transcriptome with Single-Molecule Sensitivity in Single Cells , 2010, Science.

[6]  Marco Zanella,et al.  Biological applications of gold nanoparticles. , 2008, Chemical Society reviews.

[7]  G. Whitesides,et al.  Fabrication of microfluidic systems in poly(dimethylsiloxane) , 2000, Electrophoresis.

[8]  David Piwnica-Worms,et al.  Current state of imaging protein-protein interactions in vivo with genetically encoded reporters. , 2007, Annual review of biomedical engineering.

[9]  Paul R Selvin,et al.  New fluorescent tools for watching nanometer-scale conformational changes of single molecules. , 2007, Annual review of biophysics and biomolecular structure.

[10]  Wei Sun,et al.  Resolving rotational motions of nano-objects in engineered environments and live cells with gold nanorods and differential interference contrast microscopy. , 2010, Journal of the American Chemical Society.

[11]  C. Cotman,et al.  A microfluidic culture platform for CNS axonal injury, regeneration and transport , 2005, Nature Methods.

[12]  Michael J Rust,et al.  Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM) , 2006, Nature Methods.

[13]  Wei Sun,et al.  Wavelength-dependent differential interference contrast microscopy: selectively imaging nanoparticle probes in live cells. , 2009, Analytical chemistry.

[14]  Bo Huang,et al.  Counting Low-Copy Number Proteins in a Single Cell , 2007, Science.

[15]  J. Korlach,et al.  DNA fragment sizing by single molecule detection in submicrometer-sized closed fluidic channels. , 2002, Analytical chemistry.

[16]  Susumu Sugiyama,et al.  Versatile microfluidic total internal reflection (TIR)-based devices: application to microbeads velocity measurement and single molecule detection with upright and inverted microscope. , 2009, Lab on a chip.

[17]  Cori Bargmann,et al.  Microfluidics for in vivo imaging of neuronal and behavioral activity in Caenorhabditis elegans , 2007, Nature Methods.

[18]  M. Sheetz,et al.  Tracking kinesin-driven movements with nanometre-scale precision , 1988, Nature.

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

[20]  Wei Sun,et al.  Optical imaging of non-fluorescent nanoparticle probes in live cells. , 2010, The Analyst.

[21]  Wei Sun,et al.  Influence of gold nanorod geometry on optical response. , 2010, ACS nano.

[22]  B. Cui,et al.  Optically resolving individual microtubules in live axons. , 2009, Structure.

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

[24]  Michael D. Mason,et al.  Ultra-high resolution imaging by fluorescence photoactivation localization microscopy. , 2006, Biophysical journal.

[25]  N. Jeon,et al.  Microfluidic culture platform for neuroscience research , 2006, Nature Protocols.

[26]  Piero R Bianco,et al.  Laminar flow cells for single-molecule studies of DNA-protein interactions , 2008, Nature Methods.

[27]  Wei Sun,et al.  Autocalibrated scanning-angle prism-type total internal reflection fluorescence microscopy for nanometer-precision axial position determination. , 2010, Analytical chemistry.

[28]  M. Heilemann,et al.  Subdiffraction-resolution fluorescence imaging with conventional fluorescent probes. , 2008, Angewandte Chemie.

[29]  C. Joo,et al.  Advances in single-molecule fluorescence methods for molecular biology. , 2008, Annual review of biochemistry.

[30]  Daniel T Chiu,et al.  A microfluidic model for single-cell capillary obstruction by Plasmodium falciparum-infected erythrocytes , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[31]  Kwanghun Chung,et al.  Microfluidics-enabled phenotyping, imaging, and screening of multicellular organisms. , 2010, Lab on a chip.

[32]  S. Quake,et al.  Monolithic microfabricated valves and pumps by multilayer soft lithography. , 2000, Science.

[33]  J. Lippincott-Schwartz,et al.  Imaging Intracellular Fluorescent Proteins at Nanometer Resolution , 2006, Science.

[34]  N. Friedman,et al.  Stochastic protein expression in individual cells at the single molecule level , 2006, Nature.

[35]  Mark Bates,et al.  Multicolor Super-Resolution Imaging with Photo-Switchable Fluorescent Probes , 2007, Science.

[36]  S. Gambhir,et al.  Quantum Dots for Live Cells, in Vivo Imaging, and Diagnostics , 2005, Science.

[37]  Xiaolin Nan,et al.  Organelle tracking in a living cell with microsecond time resolution and nanometer spatial precision. , 2008, Chemphyschem : a European journal of chemical physics and physical chemistry.

[38]  M. Chalfie,et al.  Green fluorescent protein as a marker for gene expression. , 1994, Science.

[39]  S. Hell.  Microscopy and its focal switch , 2008, Nature Methods.

[40]  Bin Wang,et al.  A microfluidic approach for investigating the temperature dependence of biomolecular activity with single-molecule resolution. , 2011, Lab on a chip.

[41]  R. Tsien,et al.  green fluorescent protein , 2020, Catalysis from A to Z.