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Optimization of the optical transparency of rodent tissues by modified PACT-based passive clearing

 Jiwon Woo  ;  Mirae Lee  ;  Jeong Min Seo  ;  Hyo Suk Park  ;  Yong Eun Cho 
 Experimental and Molecular Medicine, Vol.48(12) : 274, 2016 
Journal Title
 Experimental and Molecular Medicine 
Issue Date
Animals ; Bone and Bones/anatomy & histology ; Cells, Cultured ; Central Nervous System/anatomy & histology ; Embryo, Mammalian/anatomy & histology ; Guinea Pigs ; Imaging, Three-Dimensional/methods* ; Immunohistochemistry/methods* ; Light ; Lipids/isolation & purification* ; Male ; Mice, Inbred C57BL ; Microscopy, Confocal/methods* ; Models, Anatomic ; Rats
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.
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1. College of Medicine (의과대학) > Dept. of Neurosurgery (신경외과학교실) > 1. Journal Papers
Yonsei Authors
Cho, Yong Eun(조용은) ORCID logo https://orcid.org/0000-0001-9815-2720
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