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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 16, Iss. 6 — Mar. 17, 2008
  • pp: 4322–4329

Calibrating Differential Interference Contrast Microscopy with dual-focus Fluorescence Correlation Spectroscopy

Claus B. Müller, Kerstin Weiß, Walter Richtering, Anastasia Loman, and Jörg Enderlein  »View Author Affiliations


Optics Express, Vol. 16, Issue 6, pp. 4322-4329 (2008)
http://dx.doi.org/10.1364/OE.16.004322


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Abstract

We present a novel calibration technique for determining the shear distance of a Nomarski Differential Interference Contrast prism, which is used in Differential Interference Contrast microscopy as well as for the recently developed dual-focus fluorescence correlation spectroscopy. In both applications, an exact knowledge of the shear distance induced by the Nomarski prism is important for a quantitative data evaluation. In Differential Interference Contrast microscopy, the shear distance determines the spatial resolution of imaging, in dual-focus fluorescence correlation spectroscopy, it represents the extrinsic length scale for determining diffusion coefficients. The presented calibration technique is itself based on a combination of fluorescence correlation spectroscopy and dynamic light scattering. The method is easy to implement and allows for determining the shear distance with nanometer accuracy.

© 2008 Optical Society of America

OCIS Codes
(170.6280) Medical optics and biotechnology : Spectroscopy, fluorescence and luminescence
(180.1790) Microscopy : Confocal microscopy
(300.2530) Spectroscopy : Fluorescence, laser-induced

ToC Category:
Microscopy

History
Original Manuscript: January 14, 2008
Revised Manuscript: March 6, 2008
Manuscript Accepted: March 11, 2008
Published: March 14, 2008

Virtual Issues
Vol. 3, Iss. 4 Virtual Journal for Biomedical Optics

Citation
Claus B. Müller, Kerstin Weiß, Walter Richtering, Anastasia Loman, and Joerg Enderlein, "Calibrating Differential Interference Contrast Microscopy with dual-focus Fluorescence Correlation Spectroscopy," Opt. Express 16, 4322-4329 (2008)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-16-6-4322


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References

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