Effect of excitation wavelength on penetration depth in nonlinear optical microscopy of turbid media.

Effect of excitation wavelength on penetration depth in nonlinear optical microscopy of turbid media.
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DOI:
10.1117/1.3081544
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发表时间:
2009-01
影响因子:
3.5
通讯作者:
Tromberg BJ
Tromberg BJ
中科院分区:
医学3区
文献类型:
--
作者:
Balu M;Baldacchini T;Carter J;Krasieva TB;Zadoyan R;Tromberg BJ

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我们提出了一个比较研究的双光子激发荧光(TPEF)和二次谐波产生(SHG)成像在混浊介质在800和1300 nm的激发。TPEF和SHG信号在混浊组织模型中的深度依赖性衰减被用来估计光散射对每个波长处的激发强度的影响。使用1300 nm激发观察到散射长度增加50%至80%,而两种源的峰值TPEF发射强度均在表面以下10至20 μm处获得。在1300 nm处增加的穿透深度分别通过由明胶/微球和3-D器官型胶原-成纤维细胞培养物组成的组织模型的TPEF和SHG显微镜证实。我们的结果建立了1.3 μm激发在非线性光学显微镜的可行性。
We present a comparative study of two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) imaging in turbid media at 800- and 1300-nm excitation. The depth-dependent decay of TPEF and SHG signals in turbid tissue phantoms is used to estimate the impact of light scattering on excitation intensity at each wavelength. A 50 to 80% increase in scattering length is observed using 1300-nm excitation, while peak TPEF emission intensity is obtained 10 to 20 μm beneath the surface for both sources. The increased penetration depth at 1300 nm is confirmed by TPEF and SHG microscopy of tissue phantoms composed of gelatin/microspheres and 3-D organotypic collagen-fibroblast cultures, respectively. Our results establish the feasibility of 1.3-μm excitation in nonlinear optical microscopy.
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