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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 16, Iss. 7 — Mar. 31, 2008
  • pp: 5107–5117

Image heterogeneity correction in large-area, three-dimensional multiphoton microscopy

Vladimir A. Hovhannisyan, Ping-Jung Su, Yang-Fang Chen, and Chen Yuan Dong  »View Author Affiliations

Optics Express, Vol. 16, Issue 7, pp. 5107-5117 (2008)

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Large-area multiphoton laser scanning microscopy (LMLSM) can be applied in biology and medicine for high sensitivity and resolution tissue imaging. However, factors such as refractive index mismatch induced spherical aberration, emission/excitation absorption and scattering can result in axial intensity attenuation and lateral image heterogeneity, affecting both qualitative and quantitative image analysis. In this work, we describe an image correction algorithm to improve three-dimensional images in LMLSM. The method consists of multiplying the measured nonlinear signal by a three-dimensional correction factor, determined by the use of two-photon images of the appropriate specimens and specimen absorption and scattering properties at the excitation and emission wavelengths. The proposed methodology is demonstrated in correcting multiphoton images of objects imbedded in uniform fluorescent background, lung tissue, and Drosophila larva.

© 2008 Optical Society of America

OCIS Codes
(170.5810) Medical optics and biotechnology : Scanning microscopy
(180.6900) Microscopy : Three-dimensional microscopy
(190.4180) Nonlinear optics : Multiphoton processes

ToC Category:

Original Manuscript: October 26, 2007
Revised Manuscript: December 27, 2007
Manuscript Accepted: January 3, 2008
Published: March 28, 2008

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

Vladimir A. Hovhannisyan, Ping-Jung Su, Yang-Fang Chen, and Chen Y. Dong, "Image heterogeneity correction in large-area, three-dimensional multiphoton microscopy," Opt. Express 16, 5107-5117 (2008)

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