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Title: Mapping social brain circuit in the mouse brain by serial two-photon tomography Abstract: Gaining an understanding of the brain regions and circuits that govern behavior is a central goal of systems neuroscience. Social behavior is one of the evolutionally conserved core animal behavior and pathological manifestation of social behavior can lead to several disorders including autism in humans.Identifying brain circuitry involved in social behavior will lead to better understanding of how brain process and control this important behavior. One commonly used approach towards such goal is to use immediate early genes, which increase their expression transiently upon neuronal activation, to identify brain regions activated by specific external stimuli. However, conventional detection methods such as in situ hybridization or immunohistochemistry are labor intensive and semi-quantiative, thus hard to implement to examine neuronal activation throughout the entire brain. In this talk, I will present an unbiased automated method for mapping neuronal activation in the whole mouse brain at cellular resolution. Our approach is based on the visualization of the immediate early gene c-fos by serial two-photon (STP) tomography in transgenic c-fos-GFP mice. STP tomography images the mouse brain as a series of coronal sections by combining two-photon mosaic imaging and mechanical sectioning by a built-in vibratome in an automated manner. This method allows us to examine c-fos-GFP change throughout the entire mouse brain, which helps us to systematically map out brain areas with increased c-fosGFP labeling after social behavioral stimulation.The STPtomography datasets are further processed by computational methods that detect the activated GFP-positive neurons, warp their distribution to a reference brain registered to the Allen Mouse Brain Atlas, identify activated brain regions by statistical tests, and plot the anatomical connectivity between these regions. We demonstrate the use of this method for the generation of a social behavior-evoked brain activation map representing the social brain circuitry in the mouse. This systematic analysis identified a number of new regions including agranular insular cortex as well as previously described brain regions activated during social behavior. These brain regions represent candidate nodes of a complete circuit to regulate mouse social behavior. This method opens the door to systematic screening of mouse brain circuits under normal conditions and in models of human brain disorders.