Revealing intact neuronal circuitry in centimeter-sized formalin-fixed paraffin-embedded brain.

Lin, Ya-Hui; Wang, Li-Wen; Chen, Yen-Hui; Chan, Yi-Chieh; Hu, Shang-Hsiu; Wu, Sheng-Yan; Chiang, Chi-Shiun; Huang, Guan-Jie et al. · Elife · 2024

basic_science · Level V

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Abstract

Tissue-clearing and labeling techniques have revolutionized brain-wide imaging and analysis, yet their application to clinical formalin-fixed paraffin-embedded (FFPE) blocks remains challenging. We introduce HIF-Clear, a novel method for efficiently clearing and labeling centimeter-thick FFPE specimens using elevated temperature and concentrated detergents. HIF-Clear with multi-round immunolabeling reveals neuron circuitry regulating multiple neurotransmitter systems in a whole FFPE mouse brain and is able to be used as the evaluation of disease treatment efficiency. HIF-Clear also supports expansion microscopy and can be performed on a non-sectioned 15-year-old FFPE specimen, as well as a 3-month formalin-fixed mouse brain. Thus, HIF-Clear represents a feasible approach for researching archived FFPE specimens for future neuroscientific and 3D neuropathological analyses.

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