影像科学与光化学 ›› 2021, Vol. 39 ›› Issue (3): 359-363.DOI: 10.7517/issn.1674-0475.201122

• 综述与论文 • 上一篇    下一篇

基于LSCM的线扫描方向识别方法的建立与应用

刘双双, 肖桂凤, 洪晓黎, 沈逸   

  1. 浙江大学医学院, 浙江 杭州 310058
  • 收稿日期:2020-11-19 出版日期:2021-05-15 发布日期:2021-05-17
  • 通讯作者: 沈逸
  • 基金资助:
    浙江省基础公益研究计划项目(LGF20H090012)、浙江省教育厅一般科研项目(Y202043496)和中央高校基本科研业务费专项资金(2019FZA7004)资助

Establishment and Application of Line Scan Direction Recognition Method Based on LSCM

LIU Shuangshuang, XIAO Guifeng, HONG Xiaoli, SHEN Yi   

  1. School of Medicine, Zhejiang University, Hangzhou 310058, Zhejiang, P. R. China
  • Received:2020-11-19 Online:2021-05-15 Published:2021-05-17

摘要: 激光扫描共聚焦显微镜(LSCM)具有高分辨率和高光学切片的成像能力,是生命科学研究不可或缺的工具。通常采用的XY或者XYZ扫描方式成像速度慢,限制了生物研究中快速现象的追踪。本文介绍了实时快速成像的XT线扫描模式,并建立线扫描方向的识别方法。制作荧光小球样本,经过荧光小球划出不同方向的线段,使线段的一端接近荧光小球的中心而另一端远离,对线段进行XT线扫描,根据荧光信号与两端距离的远近判断扫描方向。阐述了XT线扫描在Zeiss、Nikon和Olympus共聚焦显微镜中的具体操作方法,并比较了其线扫描方向的差异。本研究建立了一套确定线扫描方向的实用方法,该方法简便易操作,具有普遍适用性,可为显微成像从业者和研究者提供借鉴。

关键词: 激光扫描共聚焦显微镜, 线扫描, 扫描方向, 荧光小球

Abstract: Laser scanning confocal microscope (LSCM) is an indispensable tool for life science research because of its high resolution and imaging ability of optical slices. The slow imaging speed of the commonly used XY and XYZ scanning methods limits the tracking of fast phenomena in biological research. This article described an XT line scanning mode for real-time fast imaging and established a method for identifying the direction of line scanning. The line segments in different directions were drawn through the fluorescent spheres, so that one end of the line segment was close to the center of the fluorescent sphere and the other end was far away. The XT line scanning was performed on the line segment, and the scanning direction was judged by the distance between the fluorescence signal and the two ends. The operating methods of XT line scanning in Zeiss, Nikon and Olympus confocal microscopes were described in detail, and the differences of the line scanning direction were compared. This method to determine the line scanning direction is simple and easy to operate, and has universal applicability, which can provide guide for application for microscopic imaging practitioners and researchers.

Key words: laser scanning confocal microscope, line scan, scan direction, fluorescent spheres