自适应光学在双光子显微成像技术中的应用  

Application of Adaptive Optics in Two⁃photon Microscopic Imaging

作　　者：章辰 高玉峰[2] 叶世蔚 李慧[2] 郑炜[2] Zhang Chen;Gao Yufeng;Ye Shiwei;Li Hui;Zheng Wei(School of Information and Communication,Shenzhen Institute of Information Technology,Shenzhen 518172,Guangdong,China;Research Center for Biomedical Optics and Molecular Imaging,Shenzhen Institutes of Advanced Technology,Chinese Academy of Sciences,Shenzhen 518055,Guangdong,China)

机构地区：[1]深圳信息职业技术学院信息与通信学院,广东深圳518172 [2]中国科学院深圳先进技术研究院生物医学光学与分子影像研究中心,广东深圳518055

出　　处：《中国激光》2023年第3期31-48,共18页Chinese Journal of Lasers

基　　金：国家自然科学基金(81927803);广东省自然科学基金(2020B121201010,2021KTSCX279);深圳市科技计划基础研究项目(RCJC20200714114433058,RCYX20210609104445093,ZDSY20130401165820357,JSGG20211029095003004);深圳信息职业技术学院博硕士科研启动扶持项目(SZIIT2022KJ043)。

摘　　要：光学显微镜是生物医学研究必不可少的工具,其中双光子显微成像技术具有大深度三维显微成像功能,被认为是深层生物组织研究的首选工具。但是,在双光子成像系统使用过程中,光学系统的装配偏差、光学元件不理想以及生物样品的不均匀性都会在成像过程中引入像差,从而降低成像质量。通过在双光子显微成像系统中引入自适应光学技术,可实现对像差的有效校正,从而提高成像的分辨率、深度和视场。介绍了双光子显微成像中的像差来源和特点,概述了自适应光学技术中不同的探测和校正方法,综述了近年来自适应光学技术在双光子显微成像中不同的应用成果,最后对自适应光学在双光子显微成像中的发展进行了展望。Significance Twophoton microscopy(TPM)has been widely used in biological imaging owing to its submicron lateral resolution,intrinsic optical sectioning,and deep penetration abilities.TPM enables the observation of cellular and subcellular dynamics in deep live tissues within highly complex and heterogeneous environments such as the mammalian brain,thereby providing critical in situ and in vivo information.However,because of the nonuniformity of the refractive index of biological tissues,the laser is distorted and scattered during propagation.Consequently,the focus point becomes a diffuse spot,which leads to a decreased imaging depth and poor resolution of TPM.Adaptive optics(AO)technology was first applied to TPM in 2000,where a genetic algorithm was used to calculate the wavefront distortion and a deformable mirror(DM)was used to correct the aberration introduced by biological samples.Since then,various AO schemes have been developed for a wide range of highresolution microscopes to advance the development of biological exploration.In this study,the sources and characteristics of aberrations in TPM are examined,and different detection and correction methods in AO are summarized.The different applications of AO in TPM in recent years are comprehensively reviewed.Progress For AO technology,wavefront detection methods are generally divided into direct wavefront detection,which uses a wavefront sensor(WS)to detect wavefronts,and indirect wavefront detection,which estimates the aberrated wavefront using iterative algorithms.In 2010,the pupilsegmentation AO method was proposed by Ji et al.This method involves the division of the pupil into several sub apertures and the use of spatial light modulator(SLM)to modulate the wavefront phase,and the imaging resolution of a fixed mouse cortex slice was restored to the neardiffractionlimited(Fig.2).In 2012,Tang et al.proposed an iterative multiphoton adaptive compensation technique that exploits the nonlinearity of multiphoton signals to determine and compensate for distortion

关 键 词：生物光学 双光子显微成像 自适应光学 波前像差 波前校正 

分 类 号：O437[机械工程—光学工程]