光片荧光显微镜与快速共聚焦显微镜用于清除小鼠脑三维成像的比较

IF 2.3 Q3 BIOCHEMICAL RESEARCH METHODS
Youngjae Ryu, Yoonju Kim, Sang-Joon Park, Sung Rae Kim, Hyung-Jun Kim, Chang Man Ha
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引用次数: 0

摘要

全脑成像通过破译组织结构、神经元回路和单个神经元追踪来理解大脑功能是很重要的。因此,已经开发了许多清除方法来获取全脑图像或三维厚组织图像。然而,成像全脑体积有几个限制,包括较长的图像采集时间,大量的数据和较长的图像后处理。基于这些限制,许多研究人员不确定哪种光学显微镜最适合成像厚组织。在这里,我们比较了快速共聚焦显微镜与光片荧光显微镜进行全脑三维成像,它们可以最快地获得图像。为了比较两种显微镜对大体积成像的影响,我们对整个小鼠大脑进行了组织清理,改变了光片荧光显微镜的样品室和低倍率物镜,并改进了样品架。我们发现,与使用4倍物镜的快速共聚焦显微镜相比,使用2.5倍物镜的光片荧光显微镜具有节省时间、大容量图像采集和高z分辨率等优点。因此,我们认为光片荧光显微镜适用于全脑成像和获得高分辨率的三维图像。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Comparison of Light-Sheet Fluorescence Microscopy and Fast-Confocal Microscopy for Three-Dimensional Imaging of Cleared Mouse Brain
Whole-brain imaging is important for understanding brain functions through deciphering tissue structures, neuronal circuits, and single-neuron tracing. Thus, many clearing methods have been developed to acquire whole-brain images or images of three-dimensional thick tissues. However, there are several limitations to imaging whole-brain volumes, including long image acquisition times, large volumes of data, and a long post-image process. Based on these limitations, many researchers are unsure about which light microscopy is most suitable for imaging thick tissues. Here, we compared fast-confocal microscopy with light-sheet fluorescence microscopy for whole-brain three-dimensional imaging, which can acquire images the fastest. To compare the two types of microscopies for large-volume imaging, we performed tissue clearing of a whole mouse brain, and changed the sample chamber and low- magnification objective lens and modified the sample holder of a light-sheet fluorescence microscope. We found out that light-sheet fluorescence microscopy using a 2.5× objective lens possesses several advantages, including saving time, large-volume image acquisitions, and high Z-resolution, over fast-confocal microscopy, which uses a 4× objective lens. Therefore, we suggest that light-sheet fluorescence microscopy is suitable for whole mouse brain imaging and for obtaining high-resolution three-dimensional images.
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来源期刊
Methods and Protocols
Methods and Protocols Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (miscellaneous)
CiteScore
3.60
自引率
0.00%
发文量
85
审稿时长
8 weeks
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