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Optimized perfusion‐based CUBIC protocol for the efficient whole‐body clearing and imaging of rat organs
Author(s) -
Matryba Paweł,
Bozycki Lukasz,
Pawłowska Monika,
Kaczmarek Leszek,
Stefaniuk Marzena
Publication year - 2018
Publication title -
journal of biophotonics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.877
H-Index - 66
eISSN - 1864-0648
pISSN - 1864-063X
DOI - 10.1002/jbio.201700248
Subject(s) - staining , biomedical engineering , clearing , perfusion , pathology , process (computing) , biology , medicine , computer science , finance , economics , operating system
Whole‐organ and whole‐body optical tissue clearing methods allowing imaging in 3 dimensions are an area of profound research interest. Originally developed to study nervous tissue, they have been successfully applied to all murine organs, yet clearing and imaging of rat peripheral organs is less advanced. Here, a modification of CUBIC clearing protocol is presented. It provides a rapid and simple approach to clear the entire adult rat organism and thus all organs within as little as 4 days. Upgraded perfusion‐based rat CUBIC protocol preserves both anatomical structure of organs and signal from proteinaceous fluorophores, and furthermore is compatible with antibody staining. Finally, it enables also volumetric cells analyses and is tailored for staining of calcium deposits within unsectioned soft tissues.