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Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics


  • Editors: Andrew Dunn and Anthony Durkin
  • Vol. 7, Iss. 2 — Feb. 1, 2012

Adaptive optics for fluorescence correlation spectroscopy

Charles-Edouard Leroux, Irène Wang, Jacques Derouard, and Antoine Delon  »View Author Affiliations

Optics Express, Vol. 19, Issue 27, pp. 26839-26849 (2011)

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Fluorescence Correlation Spectroscopy (FCS) yields measurement parameters (number of molecules, diffusion time) that characterize the concentration and kinetics of fluorescent molecules within a supposedly known observation volume. Absolute derivation of concentrations and diffusion constants therefore requires preliminary calibrations of the confocal Point Spread Function with phantom solutions under perfectly controlled environmental conditions. In this paper, we quantify the influence of optical aberrations on single photon FCS and demonstrate a simple Adaptive Optics system for aberration correction. Optical aberrations are gradually introduced by focussing the excitation laser beam at increasing depths in fluorescent solutions with various refractive indices, which leads to drastic depth-dependent bias in the estimated FCS parameters. Aberration correction with a Deformable Mirror stabilizes these parameters within a range of several tens of μm into the solution. We also demonstrate, both theoretically and experimentally, that the molecular brightness scales as the Strehl ratio squared.

© 2011 OSA

OCIS Codes
(010.1080) Atmospheric and oceanic optics : Active or adaptive optics
(170.6280) Medical optics and biotechnology : Spectroscopy, fluorescence and luminescence
(180.1790) Microscopy : Confocal microscopy

ToC Category:
Adaptive Optics

Original Manuscript: October 11, 2011
Revised Manuscript: November 25, 2011
Manuscript Accepted: December 2, 2011
Published: December 15, 2011

Virtual Issues
Vol. 7, Iss. 2 Virtual Journal for Biomedical Optics

Charles-Edouard Leroux, Irène Wang, Jacques Derouard, and Antoine Delon, "Adaptive optics for fluorescence correlation spectroscopy," Opt. Express 19, 26839-26849 (2011)

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