title: Optimization of the optical transparency of rodent tissues by modified PACT-based passive clearing.
abstract: Recently, a bio-electrochemical technique known as CLARITY was reported for three-dimensional phenotype mapping within transparent tissues, allowing clearer whole-body and organ visualization with CB-perfusion (CUBIC) and leading to the development of whole-body clearing and transparency of intact tissues with the PACT (passive clarity technique) and PARS (perfusion-assisted agent release in situ) methodologies. We evaluated the structure-function relationships in circuits of the whole central nervous system (CNS) and various internal organs using improved methods with optimized passive clarity. Thus, in the present study, we aimed to improve the original PACT procedure and passive clearing protocols for different intact rodent tissues. We determined the optimal conditions for the passive clarity method that allowed the production of a transparent whole CNS by clearing the brain and spinal cord, as well as various organs. We also improved the tissue transparency using mPACT (modified PACT), a method for direct passive clearing, and whole perfusion-based PARS-mPACT, a method for fusion clearing, and we identified the appropriate experimental conditions. These optimized methods can be used for easy and economical high-resolution mapping and phenotyping of normal and pathological elements within intact tissues.
authors.affiliation: The Spine and Spinal Cord Institute, Department of Neurosurgery, Gangnam Severance Hospital, Yonsei University, Seoul, Republic of Korea.
authors.forename: Jiwon
authors.identifier: 0000-0001-9532-7740
authors.initials: J
authors.lastname: Woo
authors.affiliation: Division of Nephrology, Department of Internal Medicine, Gangnam Severance Hospital, Yonsei University, Seoul, Republic of Korea.
authors.forename: Mirae
authors.initials: M
authors.lastname: Lee
authors.affiliation: Cellular Reprogramming and Embryo Biotechnology Laboratory and Dental Research Institute, Seoul National University School of Dentistry, Seoul, Republic of Korea.
authors.forename: Jeong Min
authors.initials: JM
authors.lastname: Seo
authors.affiliation: The Spine and Spinal Cord Institute, Department of Neurosurgery, Gangnam Severance Hospital, Yonsei University, Seoul, Republic of Korea.
authors.forename: Hyo Suk
authors.initials: HS
authors.lastname: Park
authors.affiliation: The Spine and Spinal Cord Institute, Department of Neurosurgery, Gangnam Severance Hospital, Yonsei University, Seoul, Republic of Korea.
authors.forename: Yong Eun
authors.initials: YE
authors.lastname: Cho
mesh_terms: D000818:Animals; D001842:Bone and Bones / Q000033:anatomy & histology; D002478:Cells, Cultured; D002490:Central Nervous System / Q000033:anatomy & histology; D004622:Embryo, Mammalian / Q000033:anatomy & histology; D006168:Guinea Pigs; D021621:Imaging, Three-Dimensional / Q000379:methods*; D007150:Immunohistochemistry / Q000379:methods*; D008027:Light; D008055:Lipids / Q000302:isolation & purification*; D008297:Male; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal / Q000379:methods*; D008953:Models, Anatomic; D051381:Rats
chemical_list: D008055:Lipids