A versatile method to archive and delipidate brain tissues for ensuing multiplexed immunolabeling and organ-level imaging

Lin, Y.-H.; Wang, L.-W.; Chen, Y.-H.; Chan, Y.-C.; Hu, S.-H.; Wu, S.-Y.; Chiang, C.-S.; Huang, G.-J.; Yang, S.-D.; Chu, S.-W.; Wang, K.-C.; Lin, C.-H.; Huang, P.-H.; Cheng, H.-J.; Chen, B.-C.; Chu, L.-A.

doi:10.1101/2022.11.28.518189

bioRxiv · 10.1101/2022.11.28.518189

A versatile method to archive and delipidate brain tissues for ensuing multiplexed immunolabeling and organ-level imaging

Lin, Y.-H.·Wang, L.-W.·Chen, Y.-H.·Chan, Y.-C.·Hu, S.-H.·Wu, S.-Y.·Chiang, C.-S.·Huang, G.-J.·Yang, S.-D.·Chu, S.-W.·Wang, K.-C.·Lin, C.-H.·Huang, P.-H.·Cheng, H.-J.·Chen, B.-C.·Chu, L.-A.

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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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Lin, Y.-H., Wang, L.-W., Chen, Y.-H., Chan, Y.-C., Hu, S.-H., Wu, S.-Y., Chiang, C.-S., Huang, G.-J., Yang, S.-D., Chu, S.-W., Wang, K.-C., Lin, C.-H., Huang, P.-H., Cheng, H.-J., Chen, B.-C., Chu, L.-A.. 2022-12-02. A versatile method to archive and delipidate brain tissues for ensuing multiplexed immunolabeling and organ-level imaging. https://doi.org/10.1101/2022.11.28.518189

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