留言板

尊敬的读者、作者、审稿人, 关于本刊的投稿、审稿、编辑和出版的任何问题, 您可以本页添加留言。我们将尽快给您答复。谢谢您的支持!

姓名
邮箱
手机号码
标题
留言内容
验证码

全自动推扫式高光谱显微成像系统设计与研究

唐凌宇 葛明锋 董文飞

唐凌宇, 葛明锋, 董文飞. 全自动推扫式高光谱显微成像系统设计与研究[J]. 中国光学. doi: 10.37188/CO.2021-0040
引用本文: 唐凌宇, 葛明锋, 董文飞. 全自动推扫式高光谱显微成像系统设计与研究[J]. 中国光学. doi: 10.37188/CO.2021-0040
TANG Ling-yu, GE Ming-feng, DONG Wen-fei. Design and research of fully automatic push-broom hyperspectral microscopic imaging system[J]. Chinese Optics. doi: 10.37188/CO.2021-0040
Citation: TANG Ling-yu, GE Ming-feng, DONG Wen-fei. Design and research of fully automatic push-broom hyperspectral microscopic imaging system[J]. Chinese Optics. doi: 10.37188/CO.2021-0040

全自动推扫式高光谱显微成像系统设计与研究

doi: 10.37188/CO.2021-0040
基金项目: 国家重点研发计划(No. 2017YFF0108600);中科院仪器设备研制项目(No. YJKYYQ20200038);江苏省重点研发计划(社会发展No. BE2019683);济南市“高校20条”资助项目(No. 2018GXRC016)
详细信息
    作者简介:

    唐凌宇(1997—),女,黑龙江绥化人,硕士研究生,主要从事机械工程等方面的研究。E-mail:tangly9996@163.com

    葛明锋(1987—),男,江苏南通人,博士,副研究员,硕士生导师,主要从事高光谱、荧光显微成像方面研究。E-mail:gemf@sibet.ac.cn

    董文飞(1975—),男,吉林长春人,博士,研究员,博士生导师,1996年浙江大学化学工程系化学工程专业本科毕业,1999年中国科学院长春应用化学研究所高分子物理化学专业硕士毕业,2004年获德国波兹坦大学自然科学博士学位,主要从事纳米生物医学工程及其在药物递送、在体成像和液体活检等方面的研究应用。E-mail:wenfeidong@sibet.ac.cn

  • 中图分类号: TH742

Design and research of fully automatic push-broom hyperspectral microscopic imaging system

Funds: Supported by National Key R&D Program of China (No. 2017YFF0108600); Supported by the Scientific Instrument Developing Project of the Chinese Academy of Sciences (No.YJKYYQ20200038); Primary Research & Developement Plan of Jiangsu Province(Social Development No. BE2019683); The Science and Technology Department of Jinan City (No. 2018GXRC016)
More Information
  • 摘要: 为了将光谱成像技术更方便的引入显微成像领域,本文将高光谱成像技术与显微成像技术结合,搭建出一套全自动推扫式高光谱显微成像系统。系统以倒置显微镜为主体进行设计,采用棱镜-光栅元件进行光谱分光,利用高精度二维电动运动平台进行推扫,同时结合电动对焦组件完成对焦,最后成像在高灵敏sCMOS科学相机上。根据大多数生物样本光谱检测需求,系统的光谱范围选择420~800 nm。经光谱定标和空间分辨率测试,确定系统的光谱采样率为2.06 nm,光谱分辨率均值优于3.5 nm,空间分辨率优于0.87 μm。系统引入激光自动对焦系统作为主动对焦模块,以HE染色的乳腺癌病理切片为研究对象,实验分别采用被动对焦和主动对焦方式进行推扫成像,并比较分析两种方式的优劣,认为两者均可以满足大视场成像的需求,但主动对焦成像更快速、更清晰,更加适合推扫式高光谱显微成像系统。通过对全自动推扫式高光谱显微成像系统的设计与研究,解决了高光谱显微成像中无法实时对焦的难题,实现了40倍显微物镜下3.25 mm×3.25 mm范围内全自动成像,有利于促进光谱技术在生物医学等领域中的应用。
  • 图  1  系统原理图

    Figure  1.  System schematic diagram

    图  2  棱镜-光栅分光原理图

    Figure  2.  Prism grating spectroscopic schematic diagram

    图  3  整机照片

    Figure  3.  Photos of the whole machine

    图  4  光谱定标结果

    Figure  4.  Results of spectral calibration

    图  5  分辨率板推扫图像

    Figure  5.  Push-broom image of the resolution testing board

    图  6  空间分辨率测试结果

    Figure  6.  Results of spatial resolution

    图  7  基于单帧图像清晰度评价曲线

    Figure  7.  Definition evaluation curve based on a single-frame image

    图  8  基于推扫图像的清晰度评价曲线

    Figure  8.  Definition evaluation curve based on the push-broom image

    图  9  X轴4个采样点对焦位置插值结果

    Figure  9.  Interpolation results of focusing positions of four sampling points on the X-axis

    图  10  二维平面插值结果

    Figure  10.  Interpolation results on a two-dimensional plane

    图  11  推扫路线

    Figure  11.  The route of the push-broom

    图  12  基于被动对焦的大视场推扫成像

    Figure  12.  Push-broom image with a large field of view based on passive focusing

    图  13  主动对焦原理

    Figure  13.  Principle of active focusing

    图  14  基于主动对焦的大视场推扫成像

    Figure  14.  Push-broom imaging with a large field of view based on active focusing

    表  1  单帧图像和推扫图像清晰度评价的对焦位置

    Table  1.   Focus position for clarity evaluation based on the single-frame image and the push-broom image

    Position/μm091217002500
    Single frame image/μm357376376371
    Push scan image/μm369378374367
    下载: 导出CSV

    表  2  Z值数据

    Table  2.   Z value data

    X axis/μm
    Y axis/μm
    091217002500
    0369378374371
    912378385380376
    1700385388385382
    2500367376375373
    下载: 导出CSV

    表  3  主动对焦推扫图像的对焦位置

    Table  3.   Focus position based on active focus pushbroom image

    Position /μm05001000150020002500
    Active focus position/μm369376378375371367
    Push scan image/μm369376.2377.9375.7371.1367
    下载: 导出CSV
  • [1] SORG B S, MOELLER B J, DONOVAN O, et al. Hyperspectral imaging of hemoglobin saturation in tumor microvasculature and tumor hypoxia development[J]. Journal of Biomedical Optics, 2005, 10(4): 44004. doi: 10.1117/1.2003369
    [2] LIU K X, LIN S F, ZHU S Q, et al. Hyperspectral microscopy combined with DAPI staining for the identification of hepatic carcinoma cells[J]. Biomedical Optics Express,, 2021, 12(1): 173-180. doi: 10.1364/BOE.412158
    [3] EADY M, PARK B. An unsupervised prediction model for salmonella detection with hyperspectral microscopy: a multi-year validation[J]. Applied Sciences,, 2021, 11(3): 895. doi: 10.3390/app11030895
    [4] WANG J SH, LI Q L. Quantitative analysis of liver tumors at different stages using microscopic hyperspectral imaging technology[J]. Journal of Biomedical Optics,, 2018, 23(10): 106002.
    [5] 肖功海, 舒嵘, 薛永祺. 显微高光谱成像系统的设计[J]. 光学 精密工程,2004,12(4):367-372.

    XIAO G H, SHU R, XUE Y Q. Design of microscopic hyperspectral imaging system[J]. Optics and Precision Engineering, 2004, 12(4): 367-372. (in Chinese)
    [6] 李庆利, 薛永祺, 肖功海, 等. 显微高光谱成像的生物组织定量检测机理及方法研究[J]. 科学通报,2008,53(4):493-496. doi: 10.3321/j.issn:0023-074X.2008.04.018

    LI Q L, XUE Y Q, XIAO G H, et al. Research on the mechanism and method of biological tissue quantitative detection based on micro-hyperspectral imaging[J]. Chinese Science Bulletin, 2008, 53(4): 493-496. (in Chinese) doi: 10.3321/j.issn:0023-074X.2008.04.018
    [7] ORTEGA S, GUERRA R, DÍAZ M, et al. Hyperspectral push-broom microscope development and characterization[J]. IEEE Access, 2019, 7: 122473-122491. doi: 10.1109/ACCESS.2019.2937729
    [8] ORTEGA S, FABELO H, CAMACHO R, et al. Detecting brain tumor in pathological slides using hyperspectral imaging[J]. Biomedical Optics Express,, 2018, 9(2): 818-831. doi: 10.1364/BOE.9.000818
    [9] PU H B, LIN L, SUN D W. Principles of hyperspectral microscope imaging techniques and their applications in food quality and safety detection: a review[J]. Comprehensive Reviews in Food Science and Food Safety,, 2019, 18(4): 853-866. doi: 10.1111/1541-4337.12432
    [10] SELJEBOTN S T. Continuous autofocus for line scanning hyperspectral camera[D]. Trondheim: Norwegian University of Science and Technology, 2012.
    [11] 张佳伦, 郑玉权, 蔺超, 等. 消像散的自由曲面棱镜光谱仪光学系统设计[J]. 中国光学,2020,13(4):842-851. doi: 10.37188/CO.2019-0049

    ZHANG J L, ZHENG Y Q, LIN C, et al. Design of a freeform curved prism imaging spectrometer based on an anastigmatism[J]. Chinese Optics, 2020, 13(4): 842-851. (in Chinese) doi: 10.37188/CO.2019-0049
    [12] 张天一, 朱永田, 侯永辉, 等. LAMOST高分辨率光谱仪研制[J]. 中国光学,2019,12(1):148-155. doi: 10.3788/co.20191201.0148

    ZHANG T Y, ZHU Y T, HOU Y H, et al. Construction of a LAMOST high resolution spectrograph[J]. Chinese Optics, 2019, 12(1): 148-155. (in Chinese) doi: 10.3788/co.20191201.0148
    [13] 魏巍, 崔继承, 唐玉国, 等. 医用显微成像光谱仪的光谱定标技术[J]. 光学 精密工程,2016,24(5):1015-1020. doi: 10.3788/OPE.20162405.1015

    WEI W, CUI J CH, TANG Y G, et al. Spectral calibration of medical microscopic imaging spectrometer[J]. Optics and Precision Engineering, 2016, 24(5): 1015-1020. (in Chinese) doi: 10.3788/OPE.20162405.1015
    [14] 迟明波, 韩欣欣, 徐阳, 等. 宽谱段高分辨扫描光谱定标技术[J]. 中国光学,2020,13(2):249-257. doi: 10.3788/co.20201302.0249

    CHI M B, HAN X X, XU Y, et al. Broad band and high resolution scanning spectrum calibration technology[J]. Chinese Optics, 2020, 13(2): 249-257. (in Chinese) doi: 10.3788/co.20201302.0249
  • 加载中
图(15) / 表(3)
计量
  • 文章访问数:  68
  • HTML全文浏览量:  28
  • PDF下载量:  8
  • 被引次数: 0
出版历程
  • 网络出版日期:  2021-06-02

目录

    /

    返回文章
    返回