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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

基本信息

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
项目类别：
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

项目摘要

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.