Xenium_1_2_3

Xenium In Situ Gene ExpressionSix sequential 10 um fresh frozen sections of one 34-year-old case (Table S1) of human hippocampus were placed onto a Xenium slide and processed at –20°C. The tissue was fixed and permeabilized as described in the Xenium Fixation and Permeabilization Protocol (Demonstrated protocol CG000581). A customized panel of probes targeting 316 genes were added to the tissue (Table S2). The probes were hybridized to the target RNA, ligated, and enzymatically amplified generating multiple copies for each RNA target, as described in Probe Hybridization, Ligation and Amplification user guide (User guide CG000582). The Xenium slides were, then, loaded for imaging and analysis on the Xenium Analyzer instrument, followed by decoding, according to the user guide (User guide CG000584, RRID SCR_023910). Following the analysis guidelines of 10x Genomics, negative controls spatial map was considered and valued according to the expectations as described in https://www.10xgenomics.com/support/software/xenium-onboard-analysis/latest/analysis/xoa-output-analysis-summary (10x Genomics Xenium Onboard Analysis, RRID SCR_026158). Instrument software version 1.4.2.0 and software analysis version 1.4.0.6 were used (10x Genomics Xenium Explorer, RRID SCR_025847).

Xenium原位基因表达（Xenium In Situ Gene Expression）数据集构建流程如下：选取1例34岁人类海马体样本（补充表S1）的6张连续10μm新鲜冰冻切片，置于Xenium载玻片上并于–20℃条件下完成处理。按照《Xenium固定与透化操作方案》（演示方案CG000581）对组织进行固定与透化操作。向组织中加入靶向316个基因的定制化探针组合（补充表S2）。依照《探针杂交、连接与扩增用户指南》（用户指南CG000582）所述流程，使探针与靶标RNA完成杂交、连接，并经酶促扩增为每个RNA靶标生成多个拷贝。随后将Xenium载玻片加载至Xenium分析仪设备中进行成像与分析，并依据《用户指南》（用户指南CG000584，RRID SCR_023910）完成解码步骤。遵循10x Genomics的分析规范，纳入阴性对照空间分布图，并参照https://www.10xgenomics.com/support/software/xenium-onboard-analysis/latest/analysis/xoa-output-analysis-summary 所述标准进行赋值评估（10x Genomics Xenium板载分析，RRID SCR_026158）。本实验采用的仪器软件版本为1.4.2.0，分析软件版本为1.4.0.6（10x Genomics Xenium Explorer，RRID SCR_025847）。