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.
- Revealing intact neuronal circuitry in centimeter-sized formalin-fixed paraffin-embedded brain
- Ya-Hui Lin - Natl Tsing Hua Univ, Dept Biomed Engn & Environm Sci, Hsinchu, TaiwanLi-Wen Wang - National Tsing Hua UniversityYen-Hui Chen - Acad Sinica, Inst Biomed Sci, Taipei, TaiwanYi-Chieh Chan - National Tsing Hua UniversityShang-Hsiu Hu - National Tsing Hua University, Office of Research & DevelopmentSheng-Yan Wu - Natl Tsing Hua Univ, Dept Biomed Engn & Environm Sci, Hsinchu, TaiwanChi-Shiun Chiang - National Tsing Hua University, Department of Biomedical Engineering and Environmental SciencesGuan-Jie Huang - University of TaipeiShi-Wei Chu - University of TaipeiKuo-Chuan Wang - Natl Taiwan Univ Hosp, Dept Neurosurg, Taipei, TaiwanChin-Hsien Lin - Natl Taiwan Univ Hosp, Dept Neurosurg, Taipei, TaiwanPei-Hsin Huang - Natl Taiwan Univ Hosp, Dept Pathol, Taipei, TaiwanHwai-Jong Cheng - Institute of Molecular Biology, Academia SinicaBi-Chang Chen - Institute of Cellular and Organismic Biology, Academia SinicaShang-Da Yang - National Tsing Hua University, College of Semiconductor ResearchLi-An Chu - National Tsing Hua University, Department of Biomedical Engineering and Environmental Sciences
- eLife Sciences Publications Ltd
- 28
- National Science and Technology Council in Taiwan Brain Research Center under the Higher Education Sprout Project - Ministry of Education NSTC-111-2636-B-007-007; NSTC-07-2314-B-007-003-MY3; NSTC-111-2321-B-002-016; MOST-110-2321-B-010-006 / National Science and Technology Council Brain Research Center, National Tsing-Hua University, Taiwan - National Science and Technology Council
- Journal article
- 22/05/2024
- eLife, Vol.13, RP93212
- English