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우지원Jiwon Woo

연세대학교 의과대학

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Exp. Mol. Med., Oct 02 2023 | https://doi.org/10.1038/s12276-023-01089-8 Optimization of the optical transparency of bones by PACT-based passive tissue clearing.

Byung-Ho Jin^1,2,3,12, Jiwon Woo^1,4,5,6,12, Mirae Lee^1,4,6, Seockmo Ku⁷, Hyung Seok Moon⁶, Seung Jun Ryu^1,8, Young-Min Hyun⁹, Jeong-Yoon Park1^,2,4, Sung Uk Kuh^1,2,6,10 and Yong Eun Cho^1,2,4,11

¹The Spine and Spinal Cord Institute, Department of Neurosurgery, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul 06273, Republic of Korea.

²College of Medicine, Yonsei University Graduate School, Seoul 03722, Republic of Korea.

³Department of Neurosurgery, International ST Mary´s Hospital, College of Medicine, Catholic Kwandong University, Incheon 22711, Republic of Korea.

⁴Department of Neurosurgery, Graduate School of Medical Science, Brain Korea 21 Project, Yonsei University College of Medicine, Seoul 03722, Republic of Korea.

⁵Biomedical Research Institute, Biohedron, Seoul 06230, Republic of Korea.

⁶Biomedical Research Center, Gangnam Severance Hospital, Seoul 06230, Republic of Korea.

⁷Department of Food Science and Technology, Texas A&M University, College Station, TX 77843, USA.

⁸Department of Neurosurgery, Daejeon Eulji Medical Center, Eulji University, Daejeon 35233, Republic of Korea.

⁹Department of Anatomy, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul 03722, Republic of Korea.

¹⁰Department of Medical Device Engineering and Management, Yonsei University College of Medicine, Seoul 03722, Republic of Korea.

¹¹Department of Neurosurgery, Wiltse Memorial Hospital, Suwon-si, Gyeonggi-do 16480, Republic of Korea.

¹²These authors contributed equally: Byung-Ho Jin, Jiwon Woo.

Corresponding author : Correspondence to Yong Eun Cho.

Abstract

Recent developments in tissue clearing methods such as the passive clearing technique (PACT) have allowed three-dimensional analysis of biological structures in whole, intact tissues, thereby providing a greater understanding of spatial relationships and biological circuits. Nonetheless, the issues that remain in maintaining structural integrity and preventing tissue expansion/shrinkage with rapid clearing still inhibit the wide application of these techniques in hard bone tissues, such as femurs and tibias. Here, we present an optimized PACT-based bone-clearing method, Bone-mPACT+, that protects biological structures. Bone-mPACT+ and four different decalcifying procedures were tested for their ability to improve bone tissue clearing efficiency without sacrificing optical transparency; they rendered nearly all types of bone tissues transparent. Both mouse and rat bones were nearly transparent after the clearing process. We also present a further modification, the Bone-mPACT+ Advance protocol, which is specifically optimized for processing the largest and hardest rat bones for easy clearing and imaging using established tissue clearing methods.

논문정보

형식Research article
게재일2023년 10월 (BRIC 등록일 2023-10-04)
연구진국내(교신)+국외 연구진
분야 생명과학 > 발생생물학

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