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Applied Optics

Applied Optics

APPLICATIONS-CENTERED RESEARCH IN OPTICS

  • Vol. 42, Iss. 16 — Jun. 1, 2003
  • pp: 3277–3283

Confocal total-internal-reflection fluorescence microscopy with a high-aperture parabolic mirror lens

Thomas Ruckstuhl and Stefan Seeger  »View Author Affiliations


Applied Optics, Vol. 42, Issue 16, pp. 3277-3283 (2003)
http://dx.doi.org/10.1364/AO.42.003277


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Abstract

We present a theoretical study of a new total-internal-reflection fluorescence microscope for the detection of fluorescence at a water-glass interface. The system is designed for confocal imaging and spectroscopy of nanoparticles and single molecules. Focusing and fluorescence collection through standard glass coverslips is accomplished by a parabolic mirror lens. The large aperture of the element is used to excite fluorescence within the evanescent field of a diffraction-limited focus and to collect focal emission with high efficiency. Tight focusing and supercritical excitation reduce the detection volume for fluorescent analyte molecules well below that of an attoliter (10-18 L), which can be advantageous for monitoring surface binding of single molecules without interference from fluorescence of the unbound bulk. Calculations of the electric fields in the focus region and simulated confocal imaging demonstrate the performance of the system.

© 2003 Optical Society of America

OCIS Codes
(080.3620) Geometric optics : Lens system design
(110.0180) Imaging systems : Microscopy
(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

History
Original Manuscript: September 16, 2002
Revised Manuscript: January 22, 2003
Published: June 1, 2003

Citation
Thomas Ruckstuhl and Stefan Seeger, "Confocal total-internal-reflection fluorescence microscopy with a high-aperture parabolic mirror lens," Appl. Opt. 42, 3277-3283 (2003)
http://www.opticsinfobase.org/ao/abstract.cfm?URI=ao-42-16-3277

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