Protocol for whole-tissue immunolabeling, optical clearing, and lightsheet imaging of c-Fos protein expression in unsectioned mouse brains
Summary: The c-Fos protein has been broadly utilized as a marker of neuronal activity, and conventional immunohistochemistry to determine its expression relies on tissue sections. Here, we present a protocol to visualize the endogenous c-Fos protein in intact, unsectioned mouse brains responding to...
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Elsevier
2025-06-01
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725002746 |
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| author | Koukou Fu Jing Yang |
| author_facet | Koukou Fu Jing Yang |
| author_sort | Koukou Fu |
| collection | DOAJ |
| description | Summary: The c-Fos protein has been broadly utilized as a marker of neuronal activity, and conventional immunohistochemistry to determine its expression relies on tissue sections. Here, we present a protocol to visualize the endogenous c-Fos protein in intact, unsectioned mouse brains responding to specific stimuli based on the immunolabeling-enabled three-dimensional imaging of solvent-cleared organs (iDISCO) method. We describe steps for tissue harvesting, fixation, decolorization, and permeabilization followed by whole-tissue anti-c-Fos immunolabeling. We then detail procedures for tissue embedding and optical clearing for imaging by lightsheet microscopy.For complete details on the use and execution of this protocol, please refer to Chen et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-4d9b6321c2b348b9b4ecc1cd2440bd64 |
| institution | DOAJ |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-4d9b6321c2b348b9b4ecc1cd2440bd642025-08-20T03:18:23ZengElsevierSTAR Protocols2666-16672025-06-016210386810.1016/j.xpro.2025.103868Protocol for whole-tissue immunolabeling, optical clearing, and lightsheet imaging of c-Fos protein expression in unsectioned mouse brainsKoukou Fu0Jing Yang1State Key Laboratory of Membrane Biology, School of Life Sciences, Center for Life Sciences, IDG/McGovern Institute for Brain Research, Peking University Third Hospital Cancer Center, Peking University, Beijing 100871, ChinaState Key Laboratory of Membrane Biology, School of Life Sciences, Center for Life Sciences, IDG/McGovern Institute for Brain Research, Peking University Third Hospital Cancer Center, Peking University, Beijing 100871, China; Peking Union Medical College Hospital, Beijing 100730, China; Corresponding authorSummary: The c-Fos protein has been broadly utilized as a marker of neuronal activity, and conventional immunohistochemistry to determine its expression relies on tissue sections. Here, we present a protocol to visualize the endogenous c-Fos protein in intact, unsectioned mouse brains responding to specific stimuli based on the immunolabeling-enabled three-dimensional imaging of solvent-cleared organs (iDISCO) method. We describe steps for tissue harvesting, fixation, decolorization, and permeabilization followed by whole-tissue anti-c-Fos immunolabeling. We then detail procedures for tissue embedding and optical clearing for imaging by lightsheet microscopy.For complete details on the use and execution of this protocol, please refer to Chen et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166725002746MicroscopyNeuroscienceBiotechnology and bioengineering |
| spellingShingle | Koukou Fu Jing Yang Protocol for whole-tissue immunolabeling, optical clearing, and lightsheet imaging of c-Fos protein expression in unsectioned mouse brains STAR Protocols Microscopy Neuroscience Biotechnology and bioengineering |
| title | Protocol for whole-tissue immunolabeling, optical clearing, and lightsheet imaging of c-Fos protein expression in unsectioned mouse brains |
| title_full | Protocol for whole-tissue immunolabeling, optical clearing, and lightsheet imaging of c-Fos protein expression in unsectioned mouse brains |
| title_fullStr | Protocol for whole-tissue immunolabeling, optical clearing, and lightsheet imaging of c-Fos protein expression in unsectioned mouse brains |
| title_full_unstemmed | Protocol for whole-tissue immunolabeling, optical clearing, and lightsheet imaging of c-Fos protein expression in unsectioned mouse brains |
| title_short | Protocol for whole-tissue immunolabeling, optical clearing, and lightsheet imaging of c-Fos protein expression in unsectioned mouse brains |
| title_sort | protocol for whole tissue immunolabeling optical clearing and lightsheet imaging of c fos protein expression in unsectioned mouse brains |
| topic | Microscopy Neuroscience Biotechnology and bioengineering |
| url | http://www.sciencedirect.com/science/article/pii/S2666166725002746 |
| work_keys_str_mv | AT koukoufu protocolforwholetissueimmunolabelingopticalclearingandlightsheetimagingofcfosproteinexpressioninunsectionedmousebrains AT jingyang protocolforwholetissueimmunolabelingopticalclearingandlightsheetimagingofcfosproteinexpressioninunsectionedmousebrains |