Optimization of the optical transparency of bones by PACT-based passive tissue clearing. Academic Article uri icon

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.

published proceedings

  • Exp Mol Med

author list (cited authors)

  • Jin, B., Woo, J., Lee, M., Ku, S., Moon, H. S., Ryu, S. J., ... Cho, Y. E.

complete list of authors

  • Jin, Byung-Ho||Woo, Jiwon||Lee, Mirae||Ku, Seockmo||Moon, Hyung Seok||Ryu, Seung Jun||Hyun, Young-Min||Park, Jeong-Yoon||Kuh, Sung Uk||Cho, Yong Eun

publication date

  • October 2023