Characterizing point spread functions of two-photon fluorescence microscopy in turbid medium
Characterizing point spread functions of two-photon fluorescence microscopy in turbid medium
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DOI：
10.1117/1.1578644
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发表时间：
2003-07-01
影响因子：
3.5
通讯作者：
So, P
中科院分区：
文献类型：
--
作者：
Dong, CY;Koenig, K;So, P
近年来，基于双光子激发的荧光显微镜已成为生物和生物医学成像的流行工具。其优点之一是近红外光子荧光激发可增强深度穿透，这对于深层组织成像特别有吸引力。为了充分利用双光子荧光显微镜作为生物学和医学领域的三维研究技术，表征混浊介质中的双光子成像参数非常重要。我们研究了许多散射样本中的双光子点扩散函数 (PSF)。含有 0.1-mum 荧光微球和 Liposyn III 的凝胶样品被用作模拟组织中常见的浑浊环境的模型。在由 0%、0.25%、0.5%、1% 和 2% Liposyn Ill 组成的样品中完成了水浸和油浸物镜的双光子 PSF 的完整表征。我们的结果表明，在深度达约 100 μm（油）和 200 μm（水）时，散射体（高达 2% Liposyn 111）的存在不会明显降低 PSF 宽度的目标。 (C) 2003 年光电仪器工程师协会。
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.