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KMID : 0620920230550102190
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Experimental & Molecular Medicine
2023 Volume.55 No. 10 p.2190 ~ p.2204
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Optimization of the optical transparency of bones by PACT-based passive tissue clearing
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Jin Byung-Ho
Woo Ji-Won
Lee Mi-Rae
Ku Seock-Mo
Moon Hyung-Seok
Ryu Seung-Jun
Hyun Young-Min
Park Jeong-Yoon
Kuh Sung-Uk
Cho Yong-Eun
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Abstract
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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.
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KEYWORD
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Bone, Bone development, Fluorescence imaging, Tissue engineering, Translational research
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