Department of Genetics, Evolution, Microbiology and Immunology, Institute of Biology, University of Campinas, Rua Monteiro Lobato, Campinas, Sao Paulo 13083-862, Brazil.
Graduate Program in Genetics and Molecular Biology, Institute of Biology, University of Campinas, Rua Monteiro Lobato, Campinas, Sao Paulo 13083-862, Brazil.
STAR Protoc. 2020 Oct 29;1(3):100153. doi: 10.1016/j.xpro.2020.100153. eCollection 2020 Dec 18.
This protocol combines fluorescent hybridization and immunostaining to simultaneously detect, in histological sections from the same animal, subpopulations of neurons activated after two episodes of sensory stimulation. It allows the identification of groups of cells singly activated by either stimulus or co-activated by both stimuli. Our method results in nuclear staining for mRNA and c-Fos protein, allowing better spatial and temporal resolution than previously published protocols, although it requires quick brain fixation. For complete details on the use and execution of this protocol, please refer to Carvalho et al. (2015, 2020).
本方案结合荧光杂交和免疫染色，在同一只动物的组织切片中同时检测两种感觉刺激后激活的神经元亚群。该方案可以鉴定出仅受一种刺激激活或两种刺激共同激活的细胞群。我们的方法对 mRNA 和 c-Fos 蛋白进行核染色，与以前发表的方案相比，具有更好的空间和时间分辨率，尽管它需要快速进行脑固定。如需详细了解本方案的使用和实施，请参考 Carvalho 等人（2015，2020）。
