基于贝塞尔光片照明的斑马鱼荧光成像平台。

IF 2.9 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS
Biomedical optics express Pub Date : 2025-03-27 eCollection Date: 2025-04-01 DOI:10.1364/BOE.542599
Chuhui Wang, Dongmei Su, Ziheng Zhang, Jiaju Chen, Yang Liu, Cuiyi Peng, Yachen Fan, Chenggang Yan, Sanyang Han, Minjiang Chen, Xingru Huang, Jiansong Ji, Zhenglin Chen, Dong Liu, Dongmei Yu, Peiwu Qin
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引用次数: 0

摘要

基于贝塞尔光片荧光显微镜(LSFM),建立了斑马鱼三维荧光成像平台。在三维成像过程中,激发光片保持静止,通过一个电动定位台移动样品来实现轴向扫描。为了解决三维成像中光路长度变化引起的离焦问题,在检测光路中采用了电可调透镜(ETL)。设计了一种考虑样品结构各向异性且不受加入ETL信号数学形式限制的自动重聚焦方法。结果表明,采用低数值孔径(NA)的探测物镜,ETL能获得满意的再聚焦效果。此外,还探讨了ETL对系统放大倍率和分辨率的影响。为了提高体积合成的精度,设计了一种放大倍率校准方法。该系统的设计还有利于环境噪声的记录,通过简单的背景图像减法可以提高图像质量。将这种基于硬件的背景消除方法与几种最新的荧光图像去噪算法进行了比较,结果验证了该方法的高性能。活体斑马鱼淋巴和血管结构以及血流的成像结果证明了该平台的可靠性,而无需进一步进行图像反卷积。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Zebrafish fluorescence imaging platform based on Bessel light sheet illumination.

We developed a three-dimensional (3D) zebrafish fluorescence imaging platform based on Bessel light sheet fluorescence microscopy (LSFM). During the 3D imaging process, the excitation light sheet remains static and the axial scanning is realized by moving the sample with one motorized positioning stage. To solve the defocusing problem caused by the optical path length change in 3D imaging, an electrically tunable lens (ETL) is adopted in the detection optical path. An auto-refocusing method that considers the sample structural anisotropy and has no limitation on the mathematical form of signals added to the ETL is designed. The results show that ETL can provide a satisfactory refocusing effect using detection objectives with a low numerical aperture (NA). In addition, the effects of the ETL on the system magnification and resolution are explored. A magnification calibration method is devised to refine the precision of the volume synthesis. The system design also facilitates the recording of ambient noise, which can help improve image quality with simple background image subtraction. This hardware-based background elimination method is compared with several state-of-the-art fluorescence image denoising algorithms, and the comparison results verified the high performance of this method. The imaging results of live zebrafish lymphatic and vascular structures, as well as blood flow, prove the reliability of this platform without necessitating further image deconvolution.

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来源期刊
Biomedical optics express
Biomedical optics express BIOCHEMICAL RESEARCH METHODS-OPTICS
CiteScore
6.80
自引率
11.80%
发文量
633
审稿时长
1 months
期刊介绍: The journal''s scope encompasses fundamental research, technology development, biomedical studies and clinical applications. BOEx focuses on the leading edge topics in the field, including: Tissue optics and spectroscopy Novel microscopies Optical coherence tomography Diffuse and fluorescence tomography Photoacoustic and multimodal imaging Molecular imaging and therapies Nanophotonic biosensing Optical biophysics/photobiology Microfluidic optical devices Vision research.
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