Comparison of Light-Sheet Fluorescence Microscopy and Fast-Confocal Microscopy for Three-Dimensional Imaging of Cleared Mouse Brain

Author:

Ryu Youngjae12ORCID,Kim Yoonju1,Park Sang-Joon2ORCID,Kim Sung Rae3,Kim Hyung-Jun3,Ha Chang Man1

Affiliation:

1. Research Strategy Office and Brain Research Core Facilities of Korea Brain Research Institute, Daegu 41062, Republic of Korea

2. Department of Histology, College of Veterinary Medicine, Kyungpook University, Daegu 41566, Republic of Korea

3. Dementia Research Group, Korea Brain Research Institute, Daegu 41062, Republic of Korea

Abstract

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.

Funder

Ministry of Science, ICT & Future Planning

Publisher

MDPI AG

Subject

Biochemistry, Genetics and Molecular Biology (miscellaneous),Structural Biology,Biotechnology

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