Sim, Jueun Park, Chan E. Cho, In Min, Kyeongbae Eom, Minho Han, Seungjae Jeon, Hyungju Cho, Eun-Seo Lee, Yunjeong Yun, Young Hyun Lee, Sungho Cheon, Deok-Hyeon Kim, Jihyun Kim, Museong Cho, Hyun-Ju Park, Ji-Won Kumar, Ajeet Chong, Yosep Kang, Jeong Seuk Piatkevich, Kiryl D. Jung, Erica E. Kang, Du-Seock Kwon, Seok Kyu Kim, Jinhyun Yoon, Ki-Jun Lee, Jeong-Soo Kim, Cheol-Hee Choi, Myunghwan Kim, Jin Woo Song, Mi-Ryoung Choi, Hyung Jin Boyden, Edward S. Yoon, Young-Gyu Chang, Jae-Byum 2025-02-28T08:30:27Z 2025-02-28T08:30:27Z 2025-02-21 2025-03 1936-0851 https://pubs.kist.re.kr/handle/201004/151802 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. English American Chemical Society Nanoscale Resolution Imaging of Whole Mouse Embryos Using Expansion Microscopy Article 10.1021/acsnano.4c14791 1 ACS Nano, v.19, no.8, pp.7910 - 7927 ACS Nano 19 8 7910 7927 N scie scopus 001424857900001 Chemistry, Multidisciplinary Chemistry, Physical Nanoscience & Nanotechnology Materials Science, Multidisciplinary Chemistry Science & Technology - Other Topics Materials Science Article SINGLE-CELL RESOLUTION PROTEIN SYSTEM VOLUME TISSUE BODY VISUALIZATION DISSECTION expansion microscopy super-resolution imaging whole-body imaging embryo imaging peripheralnerve system