SPECIAL SECTION ON MULTIPHOTON MICROSCOPY

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

[+] Author Affiliations
Chen-Yuan Dong

National Taiwan University, Microscopic Biophysics Laboratory, Department of Physics, Taipei?106, Taiwan

Karsten Koenig

Friedrich Schiller University Jena, Institute of Anatomy II, Teichgraben 7, D-07743?Jena, Germany

Peter So

Massachusetts Institute of Technology, Department of Mechanical Engineering, 77 Massachusetts Ave., Cambridge, Massachusetts?02139, USA E-mail: ptso@mit.edu

J. Biomed. Opt. 8(3), 450-459 (Jul 01, 2003). doi:10.1117/1.1578644
History: Received Sep. 3, 2002; Revised Mar. 14, 2003; Accepted Mar. 17, 2003; Online July 18, 2003
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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. © 2003 Society of Photo-Optical Instrumentation Engineers.

© 2003 Society of Photo-Optical Instrumentation Engineers

Citation

Chen-Yuan Dong ; Karsten Koenig and Peter So
"Characterizing point spread functions of two-photon fluorescence microscopy in turbid medium", J. Biomed. Opt. 8(3), 450-459 (Jul 01, 2003). ; http://dx.doi.org/10.1117/1.1578644


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