Labeling method for high-resolution imaging of hundreds of tissue antigens

数百种组织抗原高分辨率成像的标记方法

• 批准号: 8904759
• 负责人: JAY K TRAUTMAN
• 金额: $ 22.5万
• 依托单位: ARATOME, LLC
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8904759
• 关键词: Age; Alzheimer' s Disease; Antibodies; Antigens; Architecture; Base Sequence; Biological Neural Networks; Brain; Cataloging; Catalogs; Chemicals; Collection; Communities; Complex; Computer Simulation; DNA; Data; Data Set; Detection; Development; Dimensions; Disease; Disease Progression; Due Process; Excision; Fluorescence; Funding; Goals; Human; Image; Immunohistochemistry; Individual; Label; Lateral; Methods; Microfluidics; Molecular; Molecular Analysis; Motivation; Nerve Degeneration; Neurodegenerative Disorders; Oligonucleotides; Parkinson Disease; Pathology; Phase; Plant Resins; Population; Process; Proteins; Proteomics; Publications; Quantum Dots; Reading; Reagent; Relative (related person); Research; Research Personnel; Resolution; Running; Sampling; Services; Slice; Staining method; Stains; Synapses; System; Technology; Testing; Time; Tissue Embedding; Tissues; base; cost; crosslink; effective therapy; fluorophore; imaging modality; instrument; instrumentation; meetings; neural circuit; novel; presynaptic; protein expression; prototype; public health relevance; reconstruction; research study; screening; software development; tomography

 DESCRIPTION (provided by applicant): Neurodegenerative diseases are among the most expensive, disruptive, and least well treated of human maladies, arguably because they are not well understood. Array tomography (AT) is a new method for tissue imaging with resolution in all three dimensions sufficient to resolve individual synapses and provide quantitative characterization of multple molecular constituents. AT imaging data enable description of neural networks in the ontext of the three-­-dimensional tissue architecture. We believe such data will enable researchers to begin o comprehend the proper function of neural circuits and, importantly, to begin to undestand how it is that the various neurodegenerative processes present and progress. AT is, however, complex and expensive, and has been used in relatively few studies following the first publication in 2007 by Micheva and Smith. Aratome is currently providing AT services to the research community. The present application proposes proof-­-of-­-concept studies of a novel direct labeling method that could be the basis of a fully automated AT system, analogous to a NextGen Sequencer. The sample would be loaded into a microfluidic chamber, placed on the imaging instrument and imaging data for tens-­-tohundreds of antigens would be acquired, automatically. We would develop such an instrument, as well as the catalog of directly-­-labeled antibody and detection reagents, under Phase 2 funding.

 描述(由申请人提供)：神经退行性疾病是人类疾病中最昂贵、最具破坏性和最不受欢迎的疾病之一，可以说是因为它们没有得到很好的理解。阵列断层成像是一种新的组织成像方法，其三维分辨率足以分辨单个突触，并提供多个分子组成的定量表征。在成像数据允许描述神经网络在 在三维组织结构的背景下。我们相信这样的数据将使研究人员能够开始 O理解神经回路的正确功能，重要的是，开始了解各种神经退化过程是如何存在和发展的。然而，AT是复杂和昂贵的，在Micheva和Smith于2007年发表第一篇论文后，已被用于相对较少的研究。Aratome目前正在向研究界提供AT服务。本申请提出了一种新的直接标记方法的概念验证研究，该方法可以作为类似于NextGen测序仪的全自动AT系统的基础。样本将被加载到微流体室，放置在成像仪器上，数十到数百个抗原的成像数据将自动获取。在第二阶段的资助下，我们将开发这样一种仪器，以及直接标记的抗体和检测试剂目录。