多光子成像技术的生物医学应用新进展  

New advances in biomedical applications of multiphoton imaging technology

作　　者：李少强 耿俊娴 李艳萍 刘雄波 彭晓[1] 屈军乐[1] 刘丽炜 胡睿 Li Shao-Qiang;Geng Jun-Xian;Li Yan-Ping;Liu Xiong-Bo;Peng Xiao;Qu Jun-Le;Liu Li-Wei;Hu Rui(College of Physics and Optoelectronic Engineering,Shenzhen University,Ministry of Education/Guangdong Key Laboratory of Optoelectronic Devices and Systems,Shenzhen 518060,China)

机构地区：[1]深圳大学物理与光电工程学院,教育部/广东省光电子器件与系统重点实验室,深圳518060

出　　处：《物理学报》2020年第22期258-275,共18页Acta Physica Sinica

基　　金：国家科技部重点研发项目(批准号:2017YFA0700402);国家自然科学基金(批准号:61525503,61722508,61620106016,61835009,61935012,61961136005);深圳市自由探索项目(批准号:JCYJ20180305124902165)资助的课题。

摘　　要：多光子成像技术由于具有低侵入性、强穿透力、高空间分辨率等优点,自问世以来便成为生物医学研究的有力工具,在癌症病理、神经疾病及脑功能成像等方面取得了一系列较好的研究成果.目前,应用较为广泛的多光子成像技术是双光子激发荧光显微成像技术,其在生物医学应用中具有较大的发展潜力.本文详细阐述了多光子成像技术在多色成像、功能成像及成像深度等方面的生物医学应用新进展,包括多色双光子激发荧光显微成像、双光子激发荧光寿命显微成像、双光子光纤内窥成像和三光子显微成像技术,并简要介绍这几种多光子成像技术的原理与特性,最后展望其未来发展前景.In contrast to single photon excitation fluorescence imaging,laser scanning confocal imaging,and wide-field imaging,the multi-photon imaging has advantages of minimal invasion and deeper penetration by using nearinfrared(NIR)laser source.Moreover,it can carry out three-dimensional high-spatial-resolution imaging of biological tissues due to its natural optical tomography capability.Since its advent,multi-photon imaging has become a powerful tool in biomedicine and achieved a series of significant discoveries in cancer pathology,neurological diseases and brain functional imaging.In the past decade,as a major form of multi-photon imaging techonoogy,two-photon excited fluorescence microscopy imaging has a great potential in biomedical applications.In order to satisfy the practical biomedical applications,multi-photon imaging technologies have made significant breakthroughs in improving the deficiencies of traditional 2PEF in multi-color imaging,functional imaging,live imaging and imaging depth,such as multicolor two-photon excitation fluorescence microscopy,two-photon fluorescence lifetime imaging microscopy,two-photon fiber endoscopic imaging,and three-photon microscopy imaging technology.For example,multicolor two-photon excitation fluorescence microscopy is demonstrated to achieve simultaneous imaging of multiple fluorophores with multiple wavelenth excitation lasers or continuous spectrum.In addition,the two-photon fluorescence lifetime microscopic imaging provides a method to achieve high-resolution three-dimensional imaging of biological tissue with multidimensional information including fluorescence intensity and lifetime.In addition,two-photon optical fiber endoscopic imaging with small system size and mimal invasion is developed and used to image the tissue inside the deep organ.Finally,two-photon excitation fluorescence microscopy technique still has relatively strong scattering for brain functional imaging in vivo.Therefore,the imaging depth is limited by the signal-tobackground ratio.Three-photon microsc

关 键 词：多色双光子成像 双光子激发荧光寿命成像 双光子光纤内窥 三光子成像 

分 类 号：R318[医药卫生—生物医学工程]