Optimization of the optical transparency of bones by PACT-based passive tissue clearing.
Exp Mol Med
; 55(10): 2190-2204, 2023 10.
Article
em En
| MEDLINE
| ID: mdl-37779150
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.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Osso e Ossos
/
Imageamento Tridimensional
Idioma:
En
Ano de publicação:
2023
Tipo de documento:
Article