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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 16, Iss. 14 — Jul. 7, 2008
  • pp: 10077–10090

Single molecule correlation spectroscopy in continuous flow mixers with zero-mode waveguides

David Liao, Peter Galajda, Robert Riehn, Rob Ilic, Jason L. Puchalla, Howard G. Yu, Harold G. Craighead, and Robert H. Austin  »View Author Affiliations

Optics Express, Vol. 16, Issue 14, pp. 10077-10090 (2008)

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Zero-Mode Waveguides were first introduced for Fluorescence Correlation Spectroscopy at micromolar dye concentrations. We show that combining zero-mode waveguides with fluorescence correlation spectroscopy in a continuous flow mixer avoids the compression of the FCS signal due to fluid transport at channel velocities up to ~17 mm/s. We derive an analytic scaling relationship δ k ON k ON = δ k OFF k OFF ~ k ON + k OFF k ON 0.1 D B D F D B e SNR converting this flow velocity insensitivity to improved kinetic rate certainty in time-resolved mixing experiments. Thus zero-mode waveguides make FCS suitable for direct kinetics measurements in rapid continuous flow.

© 2008 Optical Society of America

OCIS Codes
(170.2520) Medical optics and biotechnology : Fluorescence microscopy
(230.7370) Optical devices : Waveguides
(300.2530) Spectroscopy : Fluorescence, laser-induced
(220.4241) Optical design and fabrication : Nanostructure fabrication

ToC Category:
Medical Optics and Biotechnology

Original Manuscript: March 31, 2008
Revised Manuscript: May 14, 2008
Manuscript Accepted: June 19, 2008
Published: June 23, 2008

Virtual Issues
Vol. 3, Iss. 8 Virtual Journal for Biomedical Optics

David Liao, Peter Galajda, Robert Riehn, Rob Ilic, Jason L. Puchalla, Howard G. Yu, Harold G. Craighead, and Robert H. Austin, "Single molecule correlation spectroscopy in continuous flow mixers with zero-mode waveguides," Opt. Express 16, 10077-10090 (2008)

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  1. J. B. Knight, A. Vishwanath, J. P. Brody, and R. H. Austin, "Hydrodynamic Focusing on a Silicon Chip: Mixing Nanoliters in Microseconds," Phys. Rev. Lett. 80, 3863-3866 (1998). [CrossRef]
  2. E. A. Lipman, B. Schuler, O. Bakajin, and W. A. Eaton, "Single-molecule measurement of protein folding kinetics," Science 301, 1233-1235 (2003). [CrossRef] [PubMed]
  3. D. E. Hertzog, B. Ivorra, B. Mohammadi, O. Bakajin, and J. G. Santiago, "Optimization of a microfluidic mixer for studying protein folding kinetics," Anal. Chem. 78, 4299-4306 (2006). [CrossRef] [PubMed]
  4. D. E. Hertzog, X. Michalet, M. Jäger, X. Kong, J. G. Santiago, S. Weiss, and O. Bakajin, "Femtomole mixer for microsecond kinetic studies of protein folding," Anal. Chem. 76, 7169-7178 (2004). [CrossRef] [PubMed]
  5. D. Magde, E. Elson, and W. Webb, "Thermodynamic fluctuations in a reacting system-measurement by fluorescence correlation spectroscopy," Phys. Rev. Lett. 29, 705-708 (1972). [CrossRef]
  6. E. L. Elson and D. Magde, "Fluorescence correlation spectroscopy. I. Conceptual basis and theory," Biopolymers 13, 1-27 (1974). [CrossRef]
  7. D. Magde, E. L. Elson, and W. W. Webb, "Fluorescence correlation spectroscopy. II. An experimental realization," Biopolymers 13, 29-61 (1974). [CrossRef] [PubMed]
  8. M. Gösch, H. Blom, J. Holm, T. Heino, and R. Rigler, "Hydrodynamic Flow Profiling in Microchannel Structures by Single Molecule Fluorescence Correlation Spectroscopy," Anal. Chem. 72, 3260-3265 (2000). [CrossRef] [PubMed]
  9. K. K. Kuricheti, V. Buschmann, and K. D. Weston, "Application of fluorescence correlation spectroscopy for velocity imaging in microfluidic devices," Appl. Spectrosc. 58, 1180-1186 (2004). [CrossRef] [PubMed]
  10. M. Levene, J. Korlach, S. Turner, M. Foquet, H. Craighead, and W. Webb, "Zero-Mode Waveguides for Single-Molecule Analysis at High Concentrations," Science 299, 682-686 (2003). [CrossRef] [PubMed]
  11. M. Leutenegger, M. Gósch, A. Perentes, P. Hoffmann, O. J. Martin, and T. Lasser, "Confining the sampling volume for Fluorescence Correlation Spectroscopy using a sub-wavelength sized aperture," Opt. Express 14, 956-969 (2006). [CrossRef] [PubMed]
  12. J. Wenger, D. Gérard, P.-F. Lenne, H. Rigneault, J. Dintinger, T. W. Ebbesen, A. Boned, F. Conchonaud, and D. Marguet, "Dual-color fluorescence cross-correlation spectroscopy in a single nanoaperture: towards rapid multicomponent screening at high concentrations," Opt. Express 14, 12,206-12,216 (2006). [CrossRef]
  13. S. Maiti, U. Haupts, and W. W. Webb, "Fluorescence correlation spectroscopy: Diagnostics for sparse molecules," Proceedings of the National Academy of Sciences 94, 11,753-11,757 (1997). [CrossRef]
  14. J. Widengren and U. Mets, Single Molecule Detection in Solution, chap. 3, pp. 69-120 (Wiley-VCH, 2002). [CrossRef]
  15. J. Enderlein and C. Zander, Single Molecule Detection in Solution, chap. 2, pp. 21-67 (Wiley-VCH, 2002). [CrossRef]
  16. P. G. Gloersen, "Ion-beam etching," J. Vac. Sci. Technol. 12, 28-35 (1975).
  17. R. Lee, "Microfabrication by ion-beam etching," J. Vac. Sci. Technol. 16, 164-170 (1979). [CrossRef]
  18. K. Samiee, M. Foquet, L. Guo, E. Cox, and H. Craighead, "λ -Repressor Oligomerization Kinetics at High Concentrations Using Fluorescence Correlation Spectroscopy in Zero-Mode Waveguides," Biophys. J. 88, 2145-2153 (2005). [CrossRef]
  19. T. Rindzevicius, Y. Alaverdyan, B. Sepulveda, T. Pakzeh, and M. Käll, "Nanohole plasmons in optically thin gold films," J. Phys. Chem. C. 111, 1207-1212 (2007). [CrossRef]

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