自由运动秀丽隐杆线虫双光路观测系统  

Daul-optical path observation system for tracking free-moving C.elegans

作　　者：于涛源 徐熙平[1] 张少军 张宁[1] 李宏源 王晓辉[3] YU Taoyuan;XU Xiping;ZHANG Shaojun;ZHANG Ning;LI Hongyuan;WANG Xiaohui(School of Optoelectronic Engineering,Changchun University of Science and Technology,Changchun 130022,China;Changchun Yisheng Technology Co.,Ltd,Changchun 130022,China;Changchun Institute of Applied Chemistry,Chinese Academy of Sciences,Changchun 130022,China)

机构地区：[1]长春理工大学光电工程学院,吉林长春130022 [2]长春熠生科技有限公司,吉林长春130022 [3]中国科学院长春应用化学研究所,吉林长春130022

出　　处：《光学精密工程》2024年第11期1676-1685,共10页Optics and Precision Engineering

基　　金：吉林省科技厅重点研发项目(No.20210201029GX)。

摘　　要：为了解决自由运动秀丽隐杆线虫观测系统存在的视场单一、运动数据受环境影响等问题,设计了自由运动线虫双光路观测系统。该系统融合了显微成像模块和大视场成像模块,能够同时获取线虫的显微图像和培养皿图像,并通过切换荧光模块满足线虫荧光图像的获取需求。系统将显微成像模块直接装载在XYZ三轴位移平台上,可以有效避免线虫受震动影响引发的逃逸反应。最后,使用系统对USAF1951分辨率板和不同品系的线虫进行成像。实验结果显示,显微成像模块的分辨率小于2.19μm,实现了对线虫的细胞级成像,系统获取的明场显微图像清晰显示了线虫的器官组织分布和细胞轮廓,荧光图像能够清晰显示不同荧光蛋白的分布情况。大视场成像模块照度均匀、成像清晰,通过形态学算法处理图像可以采集线虫种群行为运动数据。综上所述,该系统满足不同线虫观测需求,具备较好的稳定性和数据准确性,显著提高了线虫实验的灵活性,并为不同尺度下线虫观测信息整合提供了基础。To overcome challenges related to a limited field of view and susceptibility to environmental factors impacting data accuracy,we have developed a free-moving Caenorhabditis elegans(C.elegans)dualoptical observation system.This system integrates both a microscopy imaging module and a large-field-ofview imaging module,allowing for the simultaneous capture of C.elegans microscope images and Petri dish images.By incorporating a switchable fluorescence module,the system meets the requirements for capturing C.elegans fluorescence images.Additionally,the system includes an XYZ three-axis platform to prevent vibration-triggered escape responses from C.elegans.We utilized this system to conduct imaging experiments on USAF1951 resolution plates and various C.elegans strains.The experimental results demonstrated exceptional microscopy imaging resolution of less than 2.19 mm,vividly displaying C.elegans'organs,tissues,and cell boundaries in bright-field microscopy images,along with clear imaging of various fluorescent protein distributions.The large-field-of-view module,featuring uniform illumination and a morphological algorithm,enables the collection of behavior and motion data from the C.elegans population.In summary,this system fulfills various C.elegans tracking requirements,exhibits significant stability and data accuracy,enhances the flexibility of C.elegans experiments,and lays a foundation for integrating C.elegans tracking data across different scales.

关 键 词：显微成像 双光路成像 秀丽隐杆线虫 线虫观测系统 

分 类 号：TH742[机械工程—光学工程] TP391.41[机械工程—仪器科学与技术]