非线性光学显微镜：从基础到在活体细胞成像中的应用
Nonlinear Optical Microscopy: From Fundamentals to Applications in Live Bioimaging.
作者信息
Parodi Valentina, Jacchetti Emanuela, Osellame Roberto, Cerullo Giulio, Polli Dario, Raimondi Manuela Teresa
机构信息
Department of Chemistry, Materials and Chemical Engineering "G. Natta", Politecnico di Milano, Milan, Italy.
Istituto di Fotonica e Nanotecnologie (IFN) - CNR, Milan, Italy.
出版信息
Front Bioeng Biotechnol. 2020 Oct 9;8:585363. doi: 10.3389/fbioe.2020.585363. eCollection 2020.
A recent challenge in the field of bioimaging is to image vital, thick, and complex tissues in real time and in non-invasive mode. Among the different tools available for diagnostics, nonlinear optical (NLO) multi-photon microscopy allows label-free non-destructive investigation of physio-pathological processes in live samples at sub-cellular spatial resolution, enabling to study the mechanisms underlying several cellular functions. In this review, we discuss the fundamentals of NLO microscopy and the techniques suitable for biological applications, such as two-photon excited fluorescence (TPEF), second and third harmonic generation (SHG-THG), and coherent Raman scattering (CRS). In addition, we present a few of the most recent examples of NLO imaging employed as a label-free diagnostic instrument to functionally monitor and vital biological specimens in their unperturbed state, highlighting the technological advantages of multi-modal, multi-photon NLO microscopy and the outstanding challenges in biomedical engineering applications.
生物成像领域最近面临的一项挑战是对活体、厚且复杂的组织进行实时、非侵入式成像。在可用于诊断的不同工具中，非线性光学（NLO）多光子显微镜能够在亚细胞空间分辨率下对活样本中的生理病理过程进行无标记、非破坏性研究，从而有助于研究多种细胞功能背后的机制。在本综述中，我们讨论了NLO显微镜的基本原理以及适用于生物应用的技术，如双光子激发荧光（TPEF）、二次和三次谐波产生（SHG - THG）以及相干拉曼散射（CRS）。此外，我们展示了一些NLO成像的最新实例，这些成像被用作无标记诊断仪器，以在未受干扰的状态下对重要生物标本进行功能监测，突出了多模态、多光子NLO显微镜的技术优势以及生物医学工程应用中面临的重大挑战。
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