Nanoscale Resolution Imaging of Whole Mouse Embryos Using Expansion Microscopy

Nanoscale imaging of whole vertebrates is essential for the systematic understanding of human diseases, yet this goal has not yet been achieved. Expansion microscopy (ExM) is an attractive option for accomplishing this aim; however, the expansion of even mouse embryos at mid- and late-developmental stages, which have fewer calcified body parts than adult mice, is yet to be demonstrated due to the challenges of expanding calcified tissues. Here, we introduce a state-of-the-art ExM technique, termed whole-body ExM, that utilizes cyclic digestion. This technique allows for the super-resolution, volumetric imaging of anatomical structures, proteins, and endogenous fluorescent proteins (FPs) within embryonic and neonatal mice by expanding them 4-fold. The key feature of whole-body ExM is the alternating application of two enzyme compositions repeated multiple times. Through the simple repetition of this digestion process with an increasing number of cycles, mouse embryos of various stages up to E18.5, and even neonatal mice, which display a dramatic difference in the content of calcified tissues compared to embryos, are expanded without further laborious optimization. Furthermore, the whole-body ExM's ability to retain FP signals allows the visualization of various neuronal structures in transgenic mice. Whole-body ExM could facilitate studies of molecular changes in various vertebrates.

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Publication Details

Journal
ACS Nano
Published
2025-02-18
DOI
https://doi.org/10.1021/acsnano.4c14791
Citations
11
Primary Topic
Advanced Electron Microscopy Techniques and Applications
Type
article
Field-Weighted Citation Impact
28.03

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article

Nanoscale Resolution Imaging of Whole Mouse Embryos Using Expansion Microscopy

Myunghwan Choi, Young Hyun Yun, Deok-Hyeon Cheon, Chan E Park et al.
11 citations
ACS Nano
Advanced Electron Microscopy Techniques and Applications
28.03
article

Nanoscale Resolution Imaging of Whole Mouse Embryos Using Expansion Microscopy

Myunghwan Choi, Young Hyun Yun, Deok-Hyeon Cheon, Chan E Park, Erica E. Jung, Sungho Lee, Museong Kim, Ajeet Kumar, Kiryl D. Piatkevich, Hyungju Jeon, Du‐Seock Kang, Seok‐Kyu Kwon, Yosep Chong, Jae‐Byum Chang, Young‐Gyu Yoon, Seungjae Han, Hyung Jin Choi, Jueun Sim, Ki‐Jun Yoon, Kyeongbae Min, Minho Eom, Eun‐Seo Cho, Ji Won Park, Jin Woo Kim, Jinhyun Kim, Jeong-Soo Lee, Jang‐Hee Kim, Cheol‐Hee Kim, Yunjeong Lee, Hyun-Ju Cho, In Cho, Jeong Seuk Kang, Mi-Ryoung Song, Edward S. Boyden
article en
11 citations

Abstract

Nanoscale imaging of whole vertebrates is essential for the systematic understanding of human diseases, yet this goal has not yet been achieved. Expansion microscopy (ExM) is an attractive option for accomplishing this aim; however, the expansion of even mouse embryos at mid- and late-developmental stages, which have fewer calcified body parts than adult mice, is yet to be demonstrated due to the challenges of expanding calcified tissues. Here, we introduce a state-of-the-art ExM technique, termed whole-body ExM, that utilizes cyclic digestion. This technique allows for the super-resolution, volumetric imaging of anatomical structures, proteins, and endogenous fluorescent proteins (FPs) within embryonic and neonatal mice by expanding them 4-fold. The key feature of whole-body ExM is the alternating application of two enzyme compositions repeated multiple times. Through the simple repetition of this digestion process with an increasing number of cycles, mouse embryos of various stages up to E18.5, and even neonatal mice, which display a dramatic difference in the content of calcified tissues compared to embryos, are expanded without further laborious optimization. Furthermore, the whole-body ExM's ability to retain FP signals allows the visualization of various neuronal structures in transgenic mice. Whole-body ExM could facilitate studies of molecular changes in various vertebrates.

ACS NanoVol. 19(8)
University of Science and Technology (YE), Howard Hughes Medical Institute (US), Harvard University (US), Seoul National University (KR), McGovern Institute for Brain Research (US), Korea Advanced Institute of Science and Technology (KR), Chungnam National University (KR), Korea University (KR), Harvard University Press (US), New Generation University College (ET), Westlake University (CN), University of Illinois Chicago (US), Gwangju Institute of Science and Technology (KR), Kootenay Association for Science & Technology (CA), Korea Institute of Brain Science (KR), The Catholic University of Korea Uijeongbu St. Mary's Hospital (KR), Korean Association Of Science and Technology Studies (KR), Bio-Medical Science (South Korea) (KR), Elucid Bioimaging (US), Convergence (US), Institute of Basic Medical Sciences of the Chinese Academy of Medical Sciences, Korea Institute of Science and Technology (KR), Massachusetts Institute of Technology (US), Korea Research Institute of Bioscience and Biotechnology (KR), Sungkyunkwan University (KR), Catholic University of Korea (KR), Korea University of Science and Technology (KR)
Howard Hughes Medical Institute, Good Ventures Foundation, Korea Institute of Science and Technology, Korea Health Industry Development Institute, National Research Foundation of Korea, National Institutes of Health, Samsung
Openalex Percentile: Top 0%
Advanced Electron Microscopy Techniques and Applications
28.03
DOISourceOpenAlex
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