Research Core (Amnis ImageStreamX Core)

研究核心（Amnis ImageStreamX 核心）

• 批准号: 8725465
• 负责人: Warren D Shlomchik
• 金额: $ 15.72万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-17 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8725465
• 关键词: Animals; Appointment; Autoimmune Diseases; Autoimmunity; Award; Cell Communication; Cells; Cessation of life; Collaborations; Communities; Complex; Confocal Microscopy; Consultations; DNA Sequence Rearrangement; Data; Data Analyses; Data Set; Disease; Disease Pathway; Etiology; Event; Flow Cytometry; Fluorescence; Grant; Human Resources; Image; Imagery; Immune System Diseases; Immunology; Lasers; Learning; Left; Maintenance; Measurement; Mediating; Membrane Proteins; Methods; Microscopy; Mind; Modeling; Online Systems; Pathogenesis; Persons; Phase; Protocols documentation; Publications; Publishing; Research; Research Personnel; Rheumatism; Running; Sample Size; Sampling; Schedule; Signal Transduction; Site; System; Technology; Time; Training; Training and Education; Visit; Work; Writing; base; cell type; design; detector; digital imaging; disability; experience; instrument; medical schools; meetings; operation; ranpirnase; research study; response; success; tool; two-photon

We propose to establish a new Amnis ImageStreamX Core. Microscopy can image cytological rearrangements, cell:cell interactions, protein and membrane localization, and cell signaling events. However, such methods are neither high throughput nor quantitative. Nor are they adapted for real-time signaling or facile for the separation of defined cell types from mixtures. With these limitations in mind, Yale School of Medicine (YSM) acquired through a shared instrument grant an Amnis ImageStreamX. The Amnis ImageStreamX is a multispectral imaging instrument that acquires 12 channels of cellular imagery. By collecting large numbers of images per sample and providing numerical representation of image-based features, the ImageStreamX combines the per cell information content of standard microscopy with the statistical significance afforded by large sample sizes in standard flow cytometry. The best applications for the ImageStreamX take advantage of its abilities to locate and quantitate the distribution of signals on, within or between cells. This is the only such instrument at YSM. While there have been important discoveries made with the Amnis, the instrument is clearly underutilized. This is despite an enthusiastic initial response during a trial period wherein an expert from Amnis assisted in planning experiments, running samples and analyzing data on-site for 3 days. During this short time, preliminary data were obtained for 10 projects from 10 independent investigators. Whereas this early success was achieved with an expert on-site operator, the model applied for ongoing use of the Amnis was to treat as a user-operated 4-colorfiow cytometer. This model was not self-sustaining. Key aspects of the technology and in particular the required analysis of massive data sets rendered optimal use of the Amnis more complicated than a flow cytometer and therefore less accessible. This proposal seeks to remedy this by applying lessons learned by the P30 2P Core for which a dedicated and experienced operator participates in all phases of the work including experimental planning, training of users, imaging, assistance and training in data analysis. We propose to apply the same model to the Amnis so as to augment its impact on the study of rheumatic and immunological diseases, and the pathways that mediate their pathogenesis.

我们建议建立一个新的 Amnis ImageStreamX Core。显微镜可以对细胞学重排、细胞与细胞相互作用、蛋白质和膜定位以及细胞信号传导事件进行成像。然而，此类方法既不高通量也不定量。它们也不适合实时信号发送或易于从混合物中分离特定的细胞类型。考虑到这些限制，耶鲁大学医学院 (YSM) 通过共享仪器获得了 Amnis ImageStreamX。 Amnis ImageStreamX 是一款多光谱成像仪器，可采集 12 个通道的细胞图像。通过收集每个样本的大量图像并提供基于图像的特征的数字表示，ImageStreamX 将标准显微镜的每个细胞信息内容与标准流式细胞术中大样本量提供的统计显着性结合起来。 ImageStreamX 的最佳应用利用其定位和定量细胞上、细胞内或细胞间信号分布的能力。这是 YSM 唯一的此类仪器。虽然阿姆尼斯取得了重要发现，但该仪器显然没有得到充分利用。尽管在试用期间得到了热烈的初步反响，来自 Amnis 的专家在现场协助规划实验、运行样本和分析数据 3 天。在这短短的时间内，10 位独立研究人员获得了 10 个项目的初步数据。虽然早期的成功是由专业的现场操作员实现的，但持续使用 Amnis 所采用的模型是将其视为用户操作的 4 色流式细胞仪。这种模式不能自我维持。该技术的关键方面，特别是需要对大量数据集进行分析，使得 Amnis 的最佳使用比流式细胞仪更加复杂，因此更难获得。该提案旨在通过应用 P30 2P Core 的经验教训来解决这一问题，其中一名专门且经验丰富的操作员参与工作的所有阶段，包括实验规划、用户培训、成像、数据分析方面的协助和培训。我们建议将相同的模型应用于 Amnis，以增强其对风湿病和免疫疾病以及介导其发病机制的途径的研究的影响。