Sample Sparing Chambers for Imaging of T cell Response and Function

用于 T 细胞反应和功能成像的样品备用室

• 批准号: 9094442
• 负责人: Lance C Kam
• 金额: $ 30.71万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-18 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9094442
• 关键词: Address; Area; Automobile Driving; B-Lymphocytes; Basic Science; Biological Assay; Biological Models; Blood Cells; Blood specimen; CD28 gene; CD3 Antigens; Cell Count; Cell Survival; Cell physiology; Cells; Cellular Immunology; Cellular Structures; Clinical; Collaborations; Collection; Communities; Complex; Devices; Diagnosis; Dimensions; Disease; Elastomers; Engineering; Flow Cytometry; Frequencies; Genomics; Goals; Health; Height; Human; Image; Image Analysis; Immune; Immune System Diseases; Immune response; Immunology; Injection of therapeutic agent; Magnetism; Methods; Microfluidics; Microscopy; Modeling; Molds; Molecular; Molecular Immunology; National Institute of Allergy and Infectious Disease; Patients; Preparation; Production; Property; Proteomics; Recovery; Regulatory T-Lymphocyte; Research; Sampling; Slide; Specific qualifier value; Specimen; Surface; System; T cell response; T-Lymphocyte; Testing; Time; Tissue Sample; Translations; Work; base; cellular imaging; design; flexibility; immune function; insight; interest; lithography; polydimethylsiloxane; prototype; response; scale up; tool

 DESCRIPTION (provided by applicant): Microscopy-based assays of cell function provide important insights into human immunology and disease that are not attainable through other forms of analysis. Importantly, contemporary microscopy platforms are well suited for providing detailed spatial and functional information from small numbers of cells, a strategic property when the cells of interest are of limited frequency or availability. Unfortunately, loss of cells i a sample to dead volume spaces during sample preparation and handling, particularly when engineered surfaces are used to test cell function, severely limits the realization of this potentil. To address this low efficiency of cell analysis, we seek to develop two complementary microscopy slide chambers that allow the use of complex model systems for cell stimulation while increasing the efficiency of cell loading. The first system incorporates a modified well design that focuses cell seeding onto a small substrate area on which cell imaging and analysis takes place. The second system uses magnetic capture to localize cell seeding onto a similarly specified area, and is well suited for systems in which positive selection is used to prepare cells The goal of this project is to transition from the flexible but low-throughput methods used to fabricate the initial prototypes to higher-throughput systems. Specifically, we will transition fro elastomer-based soft lithography to injection molding of plastics commonly used in conventional labware. As a specific application of these chambers, we will examine the ability of regulatory T cells to respond to the microscale organization of TCR/CD28/LFA-1 stimulation. Successful completion of the proposed project will provide a new assay platform that is well suited for cellular and molecular studies of immune function. Strategic deployment of these systems throughout the research community will greatly accelerate the translation of basic research into clearer understanding of immune-based diseases.

 描述（由申请人提供）：基于显微镜的细胞功能测定为人类免疫学和疾病提供了通过其他形式的分析无法获得的重要见解。重要的是，当代显微镜平台非常适合提供少量细胞的详细空间和功能信息，当感兴趣的细胞频率或可用性有限时，这是一种战略特性。不幸的是，在样品制备和处理过程中，样品中的细胞损失到死体积空间，特别是当使用工程表面来测试细胞功能时，严重限制了这种潜力的实现。为了解决细胞分析效率低的问题，我们寻求开发两个互补的显微镜载玻片室，允许使用复杂的模型系统进行细胞刺激，同时提高细胞加载的效率。第一个系统采用了改进的孔设计，将细胞接种集中到进行细胞成像和分析的小基板区域上。第二个系统使用磁性捕获将细胞接种定位到类似的指定区域，并且非常适合使用正选择来制备细胞的系统。该项目的目标是从用于制造初始原型的灵活但低通量的方法过渡到更高通量的系统。具体来说，我们将从基于弹性体的软光刻过渡到传统实验室器具中常用的塑料注射成型。作为这些室的具体应用，我们将检查调节性 T 细胞对 TCR/CD28/LFA-1 刺激的微观组织作出反应的能力。 该项目的成功完成将提供一个新的检测平台，非常适合免疫功能的细胞和分子研究。这些系统在整个研究界的战略部署将大大加速基础研究转化为对免疫疾病的更清晰的理解。