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- Title
Revealing intact neuronal circuitry in centimeter-sized formalin-fixed paraffinembedded brain.
- Authors
Ya-Hui Lin; Li-Wen Wang; Yen-Hui Chen; Yi-Chieh Chan; Shang-Hsiu Hu; Sheng-Yan Wu; Chi-Shiun Chiang; Guan-Jie Huang; Shang-Da Yang; Shi-Wei Chu; Kuo-Chuan Wang; Chin-Hsien Lin; Pei-Hsin Huang; Hwai-Jong Cheng; Bi-Chang Chen; Li-An Chu
- 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 centimeterthick 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.
- Subjects
NEURAL circuitry; EXPANSION microscopy; HIGH temperatures; IMAGE analysis; THERAPEUTICS; NEUROTRANSMITTER receptors
- Publication
eLife, 2024, p1
- ISSN
2050-084X
- Publication type
Article
- DOI
10.7554/eLife.93212