Fu Koukou, Yang Jing
State 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.
State 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.
STAR Protoc. 2025 Jun 20;6(2):103868. doi: 10.1016/j.xpro.2025.103868. Epub 2025 May 31.
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..
c-Fos蛋白已被广泛用作神经元活动的标志物，而通过传统免疫组织化学来确定其表达依赖于组织切片。在此，我们提出一种基于溶剂清除器官免疫标记启用的三维成像（iDISCO）方法，用于在完整、未切片的小鼠大脑中可视化内源性c-Fos蛋白对特定刺激的反应。我们描述了组织采集、固定、脱色和通透处理的步骤，随后进行全组织抗c-Fos免疫标记。然后，我们详细说明了组织包埋和光学清除以便通过光片显微镜成像的程序。有关本方案使用和执行的完整详细信息，请参考Chen等人的研究。
