Laser-Based Mass Spectrometry Analysis of Single Cells

基于激光的单细胞质谱分析

• 批准号: 8831079
• 负责人: Jeremy L Norris
• 金额: $ 37.22万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-30 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8831079
• 关键词: Ablation; Address; Automation; Biological; Biomedical Research; Caliber; Cell Count; Cells; Charge; Clinical; Data; Detection; Development; Diagnostic; Disease; Environment; Fourier transform ion cyclotron resonance; Funding; Gases; Genomics; Health; Heterogeneity; Histology; Human; Human Biology; Human body; Image; Individual; Ions; Islet Cell; Islets of Langerhans; Knowledge; Label; Laboratories; Lasers; Lipids; Mass Spectrum Analysis; Measurement; Measures; Microdissection; Molecular; Molecular Profiling; Nature; Optics; Organ; Pancreas; Performance; Phase; Play; Population; Proteomics; Relative (related person); Research; Resolution; Resources; Role; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; System; Techniques; Technology; Therapeutic; Time; Tissues; United States National Institutes of Health; Universities; Variant; base; cell type; data acquisition; diabetic; human disease; human tissue; innovation; insight; instrument; instrumentation; mass analyzer; mass spectrometer; meetings; metabolomics; molecular phenotype; novel; public health relevance; single cell analysis

 DESCRIPTION: The proposed research seeks to describe the molecular phenotype of single cells in cell populations (e.g., organs and tissues) through the direct measurement of the lipidome and metabolome at cellular spatial resolution using MALDI mass spectrometry. Recent innovations in instrumentation conceived and built at Vanderbilt University (VU) uniquely provide the sensitivity, spatial resolution, and throughput required to meet this analytical challenge. We have developed two instrumental approaches that enable single cell analysis by MS. First, advanced laser optical systems developed in our laboratory permit targeting and ablation of features as small as a single cell with subsequent MS of the analytes using a time-of-flight mass spectrometer. This technology has sufficient spatial resolution to target single human cells in tissue without microdissection; however, sensitivity is limited due to the minimal amount of ablated material. Second, to address the need for greater sensitivity, we will employ a technique called continuous accumulation of selected ions (CASI) in conjunction with an FT-ICR mass analyzer. CASI utilizes gas-phase ion enrichment where a single mass-to- charge (m/z) range is isolated and stored in a linear ion trap prior to detection, increasing sensitivity up to 1000-fold and providing the sensitivity to measure a variety of analytes from single cells. We propose to integrate these two instrumental advances on a single instrument platform and will validate this novel platform specifically for the analysis of single cells isolated from in pancreatic islets and through the direct analysis of single cell directly from pancreas tissue sections.

 描述：这项拟议的研究试图通过使用MALDI质谱仪在细胞空间分辨率上直接测量脂体和代谢组来描述细胞群体(例如器官和组织)中单个细胞的分子表型。范德比尔特大学(VU)最近构思和制造的仪器创新独一无二地提供了满足这一分析挑战所需的灵敏度、空间分辨率和吞吐量。我们已经开发了两种仪器方法，使MS能够进行单细胞分析。第一，我们实验室开发的先进激光光学系统允许使用飞行时间质谱仪对小到单个细胞的特征进行定向和消融，随后使用飞行时间质谱仪对分析物进行MS分析。这项技术具有足够的空间分辨率，可以在不进行显微解剖的情况下靶向组织中的单个人类细胞；然而，由于消融材料的数量很少，因此灵敏度受到限制。其次，为了满足对更高灵敏度的需求，我们将结合FT-ICR质量分析仪使用一种称为连续累积选择离子(CASI)的技术。CASI利用气相离子富集法，在检测前将单个质量电荷比(m/z)范围隔离并存储在线性离子陷阱中，将灵敏度提高1000倍。 并提供了从单个细胞测量各种分析物的灵敏度。我们建议在一个仪器平台上集成这两个仪器的进步，并将验证这个新的平台，专门用于分析从胰岛和 通过直接从胰腺组织切片中直接分析单个细胞。