Abstract
Current tissue clearing and labeling approaches require freshly prepared samples to avoid problems related to over-fixation, such as loss of antigenicity or difficulty of tissue clearing. Thus, the predicament of specimens with long-term preservation limits the application of state-of-art technology to archived human brain tissues. Here, we present the development and validation of multiplex labeling of centimeter-sized archived tissue (MOCAT)—a formalin-fixed paraffin-embedding (FFPE)–based sample preservation and delipidation technique for centimeter-level tissue clearing and immunolabeling. We demonstrate that MOCAT sufficiently delipidates whole mouse brain and human brain blocks and is superior to 10% formalin for long-term antigenicity preservation; moreover, the spatial information of biomarkers offered by MOCAT-processed intact mouse brains is identical to that offered by freshly prepared SDS-delipidated intact mouse brains. MOCAT not only satisfies the need for long-term tissue storage before performing volumetric multiplex imaging for research purposes but also has the potential to be used in human clinical examination.
Competing Interest Statement
The authors have declared no competing interest.
