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

被引:78
作者
Dong, CY [1 ]
Koenig, K
So, P
机构
[1] Natl Taiwan Univ, Dept Phys, Microscop Biophys Lab, Taipei 106, Taiwan
[2] Univ Jena, Inst Anat 2, D-07743 Jena, Germany
[3] MIT, Dept Mech Engn, Cambridge, MA 02139 USA
关键词
fluorescence; microscopy; near infrared; medicine;
D O I
10.1117/1.1578644
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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-mum 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 Ill. Our results show that up to depths of about 100 (oil) and 200 mum (water), the presence of scatterers (up to 2% Liposyn 111) does not appreciably degrade the PSF widths of the objectives. (C) 2003 Society of Photo-Optical Instrumentation Engineers.
引用
收藏
页码:450 / 459
页数:10
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