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Optics Letters

| RAPID, SHORT PUBLICATIONS ON THE LATEST IN OPTICAL DISCOVERIES

  • Vol. 29, Iss. 6 — Mar. 15, 2004
  • pp: 569–571

Attoliter detection volumes by confocal total-internal-reflection fluorescence microscopy

Thomas Ruckstuhl and Stefan Seeger  »View Author Affiliations


Optics Letters, Vol. 29, Issue 6, pp. 569-571 (2004)
http://dx.doi.org/10.1364/OL.29.000569


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Abstract

We report a confocal total-internal-reflection fluorescence (TIRF) microscope that generates a detection volume for analyte molecules of less than 5 al (5×10-18>l) at a water–glass interface. Compared with conventional confocal microscopy, this represents a reduction of almost 2 orders of magnitude, which is important in isolating individual molecules at high analyte concentrations, where many biologically relevant processes occur. Diffraction-limited supercritical focusing and fluorescence collection is accomplished by a parabolic mirror objective. The system delivers TIRF images with excellent spatial resolution and detects single molecules with a high signal-to-background ratio.

© 2004 Optical Society of America

OCIS Codes
(170.6280) Medical optics and biotechnology : Spectroscopy, fluorescence and luminescence
(180.1790) Microscopy : Confocal microscopy
(180.2520) Microscopy : Fluorescence microscopy
(190.4350) Nonlinear optics : Nonlinear optics at surfaces
(260.6970) Physical optics : Total internal reflection

Citation
Thomas Ruckstuhl and Stefan Seeger, "Attoliter detection volumes by confocal total-internal-reflection fluorescence microscopy," Opt. Lett. 29, 569-571 (2004)
http://www.opticsinfobase.org/ol/abstract.cfm?URI=ol-29-6-569


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References

  1. S. Nie, D. T. Chiu, and R. N. Zare, Science 266, 1018 (1994).
  2. S. T. Hess, S. Huang, A. A. Heikal, and W. W. Webb, Biochemistry 41, 697 (2002).
  3. M. J. Levene, J. Korlach, S. W. Turner, M. Foquet, H. G. Craighead, and W. W. Webb, Science 299, 682 (2003).
  4. A. D. Stout and D. Axelrod, Appl. Opt. 28, 5237 (1989).
  5. T. E. Starr and N. L. Thompson, J. Phys. Chem. B 106, 2365 (2002).
  6. T. Ruckstuhl and S. Seeger, Appl. Opt. 42, 3277 (2003).
  7. V. Buschmann, K. D. Weston, and M. Sauer, Bioconjugate Chem. 14, 195 (2003).
  8. M. Rankl, S. Laib, and S. Seeger, Colloids Surf. B 30, 177 (2003).
  9. E. Bismuto, E. Gratton, and D. C. Lamb, Biophys. J. 81, 3510 (2001).
  10. D. A. Koppel, Phys. Rev. A 10, 1938 (1974).
  11. T. E. Starr and N. L. Thompson, Biophys. J. 80, 1575 (2001).

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