摘要
提出一种非轴向扫描的细胞显微深度成像技术,在显微系统中加入菲涅耳透镜,利用菲涅耳透镜的色散将不同激发光波长聚焦到不同的轴向位置,以实现对两个或多个焦平面同时成像.基于405nm和532nm两种激发光波长,在传统的荧光显微镜的激发路径中加入对应的两个成像探测器来探测两个不同焦平面所对应像面的成像信息,搭建得到一个能够实现探测深度约为12μm的基于菲涅耳透镜的荧光显微深度成像系统,并与基于显微物镜色差无菲涅耳透镜的荧光显微深度成像系统的成像深度和轴向分辨率进行实验对比.实验结果表明:加入菲涅耳透镜能够实现系统对不同焦面的同时成像;对于同一荧光波段,保证系统横向分辨率的同时,扩大了成像景深.该系统可以实现荧光生物细胞内部不同深度处的多波段同时探测.
A technique about non-axial-scanning microscopic cell depth imaging is proposed.Fresnel lens are added into the microscopic system,and the chromatic aberration of a Fresnel lens is exploited to focus different excitation light wavelengths into different axial positions.It can realize the imaging of two or more focal planes.Based on the excitation light wavelengths of 405 nm and 532 nm,two imaging detectors are added into the excitation path of a conventional fluorescence microscope to detect the imaging information of the image plane corresponding to two different focal planes.A fluorescence microscopic depth imaging system with a detection depth of about 12μm is established.The imaging depth and axial resolution of the proposed system are compared with those of a fluorescence microscopic depth imaging system based on a microscope-less Fresnel lens.Experimental results show that the addition of Fresnel lens enables simultaneous imaging of different focal planes.For the same fluorescence band,the lateral resolution of the system is ensured while the imaging depth of field is expanded,the system can achieve multi-band simultaneous detection at different depths of fluorescent biological cells.
作者
黄新荣
郑继红
刘悠嵘
朱天赟
缪涛
HUANG Xin-rong;ZHENG Ji-hong;LIU You-rong;ZHU Tian-yun;MIAO Tao(Engineering Research Center of Optical Instrument and System,Ministry of Education,Shanghai Key Lab of Modern Optical System,School of Optical-Electrical and Computer Engineering, University of Shanghai for Science and Technology,Shanghai 200093,China)
出处
《光子学报》
EI
CAS
CSCD
北大核心
2018年第9期255-261,共7页
Acta Photonica Sinica
基金
上海市科学技术委员会科研计划项目(No.18060502500)资助。
关键词
显微系统
荧光显微成像
菲涅耳透镜
荧光细胞
焦距偏移
深度成像
轴向分辨率
Microscopy system
Fluorescent microscopic imaging
Fresnel lens
Fluorescent cell
Focal shift
Depth imaging
Axial resolution
作者简介
黄新荣(1994-),女,硕士研究生,主要研究方向为荧光显微细胞成像系统.Email:xinrong5126@163.com;导师(通讯作者):郑继红(1975-),女,教授,博士,主要研究方向为聚合物分散液晶材料及光电器件、信息光学等.Email:jihongzheng@sina.com。
