Digital Spatial Profiler Analysis Instrument

项目总结 Sanford Burnham Prebys医学发现研究所(SBP)拥有约40个兴趣广泛的实验室，横跨 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 老年性疾病，包括神经退行性变。细胞-细胞异质性和空间分布的详细分析 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，SBP请求支持 NanoStringGeoMX数字空间剖面仪(DSP)可实现蛋白质或蛋白质的空间分辨率和数字定量 在组织切片上进行高度多重分析的mRNAs。GeoMX DSP将在SBP中架起空间分析桥梁 组织学核心(通过传统的免疫组织化学或免疫荧光)和单细胞测序 SBP基因组学核心中的方法。前者提供目标在组织中分布的空间信息 或肿瘤，但非常低的丛体，因为只有少数a靶点可以同时分析。后者是非常 高复合体，在单个细胞中检测到数千个表达的基因，但没有给出空间信息，因为它开始 用组织/肿瘤来源的细胞悬浮液。GeoMX数字信号处理器产生组织的空间信息或 肿瘤但也是非常高的复合体，检测到数十种蛋白质、1000个或RNA甚至整个转录组 同时。蛋白质或RNA定量在定义的感兴趣区域(ROI)内执行 在形态上或通过表达多达4个感兴趣的蛋白质或RNA标记。ROI被灵活地定义并且可以， 例如，是组织的离散区域或散布在组织中的细胞子集。这款多功能 仪器允许分析福尔马林固定或新鲜冰冻切片，人或鼠组织，RNA或 蛋白质分析和有针对性或无偏见的整个转录组分析。这些特点是空间分辨率高、 对于SBP的主要用户和其他用户来说，复杂分析和多功能性使GeoMX DSP成为一项变革性的技术。 该应用程序由SBP的10个主要用户支持，他们将占据仪器可用使用量的75% 时间(澳特)。与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的环境以及我们教员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.