双色荧光辐射差分超分辨显微系统研究  

Dual-color fluorescence emission difference super-resolution microscopy

作　　者：张智敏 匡翠方[1] 王子昂[1] 朱大钊 陈友华[1] 李传康 刘文杰 刘旭[1] ZHANG Zhi-min, KUANG Cui-fang , WANG Zi-ang, ZHU Da-zhao, CHEN You-hua, LI Chuan-kang, LIU Wen-jie, LIU Xu(College of Optical Science and Engineering ,Zhejiang University,Hangzhou 310058, Chin)

机构地区：[1]浙江大学光电科学与工程学院,浙江杭州310058

出　　处：《中国光学》2018年第3期329-336,共8页Chinese Optics

基　　金：国家重大科学仪器设备开发资助项目(No.2016YFF0101401)~~

摘　　要：为了拓展荧光辐射差分(Fluorescence Emission Difference,FED)显微术的应用,使得该方法可以同时对生物样品的不同组织结构进行超分辨成像,本文对双色FED显微系统展开了研究。FED的基本原理是将实心光斑扫描得到的共焦显微图像减去空心光斑扫描得到的负共焦图像,以此获得超分辨显微图像。在对单色FED显微系统进行研究后,本文提出了一种可行的双色FED显微成像系统方案。实验结果表明,在488 nm和640 nm激发光下,该系统在荧光颗粒上分别实现了135 nm和160 nm的空间分辨率,另外也能对生物样品的不同组织进行多色同时超分辨显微成像,满足了实际应用的要求。To perform super-resolution imaging of different tissue structures of biological samples using fluorescence radiation differential microscopy simultaneously,a dual-color FED microscopy system is studied in this paper.The basic principle of the FED is to remove the confocal microscopy image obtained by scanning the solid spot from the confocal microscopy image obtained by scanning the hollow spot to obtain a super-resolution microscopy image.Based on the study of the monochromatic FED microscopy system,a feasible dual-color FED microscopy imaging system is proposed and imaging experiments are performed on fluorescent particles in this paper.The experimental results indicate that under excitation light of 488 nm and 640 nm,the system realizes spatial resolution of 135 nm and 160 nm of the fluorescent particles respectively.In addition,this systemcan also perform multi-color super-resolution microscopy imaging simultaneously on different tissues of biological samples,which meets the requirements of practical applications.

关 键 词：光学显微 衍射极限 荧光 荧光辐射差分 超分辨 

分 类 号：TH742[机械工程—光学工程]