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Optics Letters

Optics Letters


  • 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)

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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

Thomas Ruckstuhl and Stefan Seeger, "Attoliter detection volumes by confocal total-internal-reflection fluorescence microscopy," Opt. Lett. 29, 569-571 (2004)

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