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Exp Mol Med. 2016 Dec 2;48(12):e274. doi: 10.1038/emm.2016.105.

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

Author information

1
The Spine and Spinal Cord Institute, Department of Neurosurgery, Gangnam Severance Hospital, Yonsei University, Seoul, Republic of Korea.
2
Division of Nephrology, Department of Internal Medicine, Gangnam Severance Hospital, Yonsei University, Seoul, Republic of Korea.
3
Cellular Reprogramming and Embryo Biotechnology Laboratory and Dental Research Institute, Seoul National University School of Dentistry, Seoul, Republic of Korea.

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.

PMID:
27909337
PMCID:
PMC5192069
DOI:
10.1038/emm.2016.105
[Indexed for MEDLINE]
Free PMC Article

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