Digital Spatial Profiler Analysis Instrument

项目摘要 Sanford Burnham Prebys医学发现研究所（SBP）拥有约40个实验室，这些实验室具有广泛的兴趣， 正常组织干细胞，组织稳态和功能，神经科学，癌症，衰老和退行性疾病 包括神经退化在内的衰老疾病。详细分析了细胞-细胞异质性和空间 跨多细胞组织的组织或解体是理解这些生理和 病理组织状态和功能。为此，为了保持领先地位，战略和业务伙伴关系请求支持 Nanostring GeoMX数字空间轮廓仪（DSP），可实现蛋白质的空间分辨数字定量， 在组织切片上的高度多重测定中的mRNA。GeoMX DSP将在SBP的空间分析中架起桥梁 组织学核心（通过传统免疫组织化学或免疫荧光）和单细胞测序 SBP的Genomics Core。前者给出了组织中靶分布的空间信息 或肿瘤，但是非常低的复杂性，因为只有少数靶可以同时分析。后者非常 高倍数，检测单个细胞中表达的1000个基因，但没有给出空间信息，因为它开始于 组织/肿瘤来源的细胞悬液。GeoMX DSP产生跨组织的空间信息， 但也非常复杂，检测几十种蛋白质，1000或RNA，甚至整个转录组 同步蛋白质或RNA定量在目标区域（ROI）内进行， 形态学上或通过表达多达4种感兴趣的蛋白质或RNA标记物。ROI定义灵活， 例如是组织的离散区域或散布在整个组织中的细胞亚群。这种多用途 仪器允许分析福尔马林固定或新鲜冷冻切片、人或小鼠组织、RNA或 蛋白质分析和靶向或无偏的全转录组分析。这些空间分辨率的特点，高 复杂的分析和多功能性使GeoMX DSP成为SBP主要用户和其他用户的变革性技术。 该应用程序由SBP的10个“主要用户”支持，他们将占用仪器可用使用量的75 时间（AUT）。像任何新的变革性技术一样，GeoMX DSP将生成以前无法获得的数据 和洞察力，导致新的假设，新的资金和合作。这将加强研究 SBP的环境和我们的教师与其他研究人员的互动和合作。

PROJECT SUMMARY Sanford Burnham Prebys Medical Discovery Institute (SBP) hosts ~40 labs with wide-ranging interests, spanning normal tissue stem cells, tissue homeostasis and function, neuroscience, cancer, aging and degenerative diseases of aging, including neurodegeneration. A detailed analysis of cell – cell heterogeneity and spatial organization or disorganization across a multicellular tissue is key to understanding these physiological and pathological tissue states and functions. To this end, and to stay cutting-edge, SBP requests support for a Nanostring GeoMX Digital Spatial Profiler (DSP) to enable spatially resolved, digital quantitation of proteins or mRNAs in a highly multiplexed assay on tissue sections. The GeoMX DSP will bridge spatial analyses in SBP's Histology Core (by traditional immunohistochemistry or immunofluorescence) and single cell sequencing approaches in SBP's Genomics Core. The former gives spatial information on target distribution across a tissue or tumor, but is very low plex in that only a handful a targets can be analyzed at the same time. The latter is very high plex, detecting 1000s of expressed genes in a single cell, but gives no spatial information because it starts with a tissue/tumor-derived cell suspension. The GeoMX DSP yields spatial information across the tissue or tumor but is also very high plex, detecting dozens of proteins, 1000s or RNAs or even whole transcriptome simultaneously. Protein or RNA quantitation is performed within regions of interest (ROI), defined morphologically or by expression of up to 4 protein or RNA markers of interest. ROI are flexibly defined and can, for example, be a discrete region of the tissue or a subset of cells scattered throughout the tissue. This versatile instrument permits analysis of either formalin-fixed or fresh-frozen sections, human or mouse tissues, RNA or protein analyses and targeted or unbiased whole transcriptome analyses. These traits of spatial resolution, high plex analysis and versatility make GeoMX DSP a transformative technology for SBP's major users and others. This application is supported by 10 “major users” at SBP who will take 75% of the instrument's available use time (AUT). Like any novel transformative technology, GeoMX DSP will generate previously unattainable data and insights leading to novel hypotheses, new funding and collaborations. This will enhance the research environment at SBP and our faculty's interactions and collaborations with other researchers.