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Journal of Biomedical Optics

Characterizing point spread functions of two-photon fluorescence microscopy in turbid medium
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Paper Abstract

In recent years, fluorescence microscopy based on two-photon excitation has become a popular tool for biological and biomedical imaging. Among its advantages is the enhanced depth penetration permitted by fluorescence excitation with the near-infrared photons, which is particularly attractive for deep-tissue imaging. To fully utilize two-photon fluorescence microscopy as a three-dimensional research technique in biology and medicine, it is important to characterize the two-photon imaging parameters in a turbid medium. We investigated the two-photon point spread functions (PSFs) in a number of scattering samples. Gel samples containing 0.1-μm fluorescent microspheres and Liposyn III were used as phantoms mimicking the turbid environment often found in tissue. A full characterization of the two-photon PSFs of a water and oil immersion objective was completed in samples composed of 0, 0.25, 0.5, 1, and 2% Liposyn III. Our results show that up to depths of about 100 (oil) and 200 µm (water), the presence of scatterers (up to 2% Liposyn III) does not appreciably degrade the PSF widths of the objectives.

Paper Details

Date Published: 1 July 2003
PDF: 10 pages
J. Biomed. Opt. 8(3) doi: 10.1117/1.1578644
Published in: Journal of Biomedical Optics Volume 8, Issue 3
Show Author Affiliations
Chen-Yuan Dong, National Taiwan Univ. (Taiwan)
Karsten Koenig, Friedrich-Schiller-Univ. Jena (Germany)
Peter T. C. So, Massachusetts Institute of Technology (United States)

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