Data_Sheet_1_High-Throughput Strategy for Profiling Sequential Section With Multiplex Staining of Mouse Brain.docx

The brain modulates specific functions in its various regions. Understanding the organization of different cells in the whole brain is crucial for investigating brain functions. Previous studies have focused on several regions and have had difficulty analyzing serial tissue samples. In this study, we introduced a pipeline to acquire anatomical and histological information quickly and efficiently from serial sections. First, we developed a serial brain-slice-staining method to stain serial sections and obtained more than 98.5% of slices with high integrity. Subsequently, using the self-developed analysis software, we registered and quantified the signals of imaged sections to the Allen Mouse Brain Common Coordinate Framework, which is compatible with multimodal images and slant section planes. Finally, we validated the pipeline with immunostaining by analyzing the activity variance in the whole brain during acute stress in aging and young mice. By removing the problems resulting from repeated manual operations, this pipeline is widely applicable to serial brain slices from multiple samples in a rapid and convenient manner, which benefits to facilitate research in life sciences.

大脑通过不同脑区调控特定功能。解析全脑不同细胞的组织构成，对于研究大脑功能至关重要。既往研究多聚焦于少数脑区，且难以开展连续组织样本的分析。本研究构建了一套可快速高效获取连续脑片解剖与组织学信息的标准化流程。首先，我们开发了连续脑片染色方法对连续切片进行染色，获得了完整性超过98.5%的脑片。随后，借助自研的分析软件，我们将成像后的脑片信号配准并定量至艾伦小鼠脑通用坐标框架（Allen Mouse Brain Common Coordinate Framework），该框架可兼容多模态成像数据与倾斜切片平面。最后，我们通过免疫染色分析年轻与衰老小鼠急性应激过程中的全脑活动差异，对该流程进行了验证。该流程解决了重复手动操作带来的诸多问题，可快速便捷地适配多样本的连续脑片分析，有助于推动生命科学领域的研究进展。