Raman Microscopy: A Noninvasive Method to Visualize the Localizations of Biomolecules in the Cornea
Raman Microscopy: A Noninvasive Method to Visualize the Localizations of Biomolecules in the Cornea
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DOI：
10.1097/ico.0000000000001369
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发表时间：
2017-11-01
期刊：
影响因子：
2.8
通讯作者：
Kano, Hideaki
中科院分区：
文献类型：
--
作者：
Kaji, Yuichi;Akiyama, Toshihiro;Kano, Hideaki
目的：在体内和原位可视化的生物分子没有预处理将是重要的诊断和治疗的眼部疾病在未来。近年来，基于分子与光子之间的非线性相互作用的多光子显微术已被应用于揭示各种分子在组织中的定位。我们的目的是使用多模态多光子显微镜可视化特定的生物分子在大鼠corneas.Methods的本地化：角膜的多光子图像获得的相干反斯托克斯拉曼散射，三阶和频产生，和二次谐波产生的非线性信号。在第三层，含粘附复合物的基底膜和Bowman层的定位清晰可见。顺序和频率生成图像。在二次谐波图像中，在角膜基质中观察到I型胶原的精细结构。在相干反斯托克斯拉曼散射images.Conclusions：成像技术取得了重大进展，并在医学领域得到了应用。光学相干断层扫描和共焦显微镜被广泛使用，但不能提供角膜分子结构的信息。相比之下，多光子显微镜提供了活组织分子结构的信息。使用这种技术，我们成功地可视化各种生物分子，包括脂质，蛋白质和核酸在角膜中的定位。我们推测，多光子显微镜将提供必要的信息，角膜的生理和病理条件，以及在组织中的分子定位，而无需预处理。
Purpose: In vivo and in situ visualization of biomolecules without pretreatment will be important for diagnosis and treatment of ocular disorders in the future. Recently, multiphoton microscopy, based on the nonlinear interactions between molecules and photons, has been applied to reveal the localizations of various molecules in tissues. We aimed to use multimodal multiphoton microscopy to visualize the localizations of specific biomolecules in rat corneas.Methods: Multiphoton images of the corneas were obtained from nonlinear signals of coherent anti-Stokes Raman scattering, third-order sum frequency generation, and second-harmonic generation.Results: The localizations of the adhesion complex-containing basement membrane and Bowman layer were clearly visible in the third-order sum frequency generation images. The fine structure of type I collagen was observed in the corneal stroma in the second-harmonic generation images. The localizations of lipids, proteins, and nucleic acids (DNA/RNA) was obtained in the coherent anti-Stokes Raman scattering images.Conclusions: Imaging technologies have progressed significantly and been applied in medical fields. Optical coherence tomography and confocal microscopy are widely used but do not provide information on the molecular structure of the cornea. By contrast, multiphoton microscopy provides information on the molecular structure of living tissues. Using this technique, we successfully visualized the localizations of various biomolecules including lipids, proteins, and nucleic acids in the cornea. We speculate that multiphoton microscopy will provide essential information on the physiological and pathological conditions of the cornea, as well as molecular localizations in tissues without pretreatment.