Research Core (Amnis ImageStreamX Core)

研究核心（Amnis ImageStreamX 核心）

• 批准号: 8444012
• 负责人: Warren D Shlomchik
• 金额: $ 9.97万
• 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/8444012
• 关键词: Autoimmune Diseases; Autoimmunity; Cell Communication; Cells; Cessation of life; Confocal Microscopy; DNA Sequence Rearrangement; Data; Data Analyses; Data Set; Disease; Disease Pathway; Etiology; Event; Flow Cytometry; Grant; Image; Imagery; Immune System Diseases; Learning; Mediating; Membrane Proteins; Methods; Microscopy; Mind; Modeling; Pathogenesis; Phase; Research; Research Personnel; Rheumatism; Running; Sample Size; Sampling; Signal Transduction; Site; Technology; Time; Training; Work; base; cell type; disability; experience; instrument; medical schools; ranpirnase; research study; response; success; tool

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核心。显微镜可以图像细胞学重排，细胞：细胞相互作用，蛋白质和膜定位以及细胞信号传导事件。但是，这种方法既不是高吞吐量也不是定量的。它们也不适用于实时信号传导或便利，以将定义的细胞类型从混合物中分离出来。考虑到这些局限性，耶鲁大学医学院（YSM）通过共享的仪器授予Amnis Imagestreamx获得。 Amnis ImagestreamX是一种多光谱成像仪器，可获得12个细胞成像通道。通过收集每个样品的大量图像并提供基于图像的特征的数值表示，ImageStreamx结合了标准显微镜的每个细胞信息含量与标准流式细胞仪中大样本量提供的统计显着性。 Imagestreamx的最佳应用利用了其能力定位和定量单元内或之间的信号分布。这是YSM唯一的仪器。虽然羊膜有重要的发现，但该仪器显然未被充分利用。尽管在试验期间有热情的初步响应，但AMNIS的专家协助计划实验，运行样品并在现场分析数据3天。在此短时间内，从10位独立研究人员那里获得了10个项目的初步数据。尽管这一早期成功是通过专家现场运营商实现的，但该模型旨在持续使用AMNIS作为用户操作的4-Colorfiow细胞图仪。这个模型不是自我维持的。技术的关键方面，尤其是对大规模数据集的所需分析，使AMNIS的最佳使用比流式细胞仪更为复杂，因此易于访问。该建议旨在通过应用P30 2P核心所学的教训来解决此问题，专门的经验丰富的操作员参与了工作的各个阶段，包括实验计划，培训用户，成像，帮助和数据分析中的培训。我们建议将相同的模型应用于AMNIS，以增强其对风湿性和免疫疾病研究的影响，以及介导其发病机理的途径。