Single Cell Analysis via Nanoscale Tip-Enhanced Laser Ablation Mass Spectrometry

通过纳米级尖端增强激光烧蚀质谱进行单细胞分析

• 批准号: 9047750
• 负责人: Kermit King Murray
• 金额: $ 59.78万
• 依托单位: ANASYS INSTRUMENTS CORPORATION
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-01 至 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9047750
• 关键词: Ablation; Address; Area; Atomic Force Microscopy; Benchmarking; Biological; Brain imaging; Cells; Charge; Chemicals; Cocaine; Collaborations; Detection; Development; Devices; Disease; Electrospray Ionization; Geometry; Goals; Image; Image Analysis; Imaging Device; Ions; Laboratories; Lasers; Lipids; Louisiana; Mass Spectrum Analysis; Methods; Modality; National Institute of Drug Abuse; Online Systems; Outcome; Peptides; Phase; Physiologic pulse; Progress Reports; Rattus; Research; Resolution; Sampling; Scanning Probe Microscopes; Small Business Technology Transfer Research; Source; Spottings; Staging; System; Technology; Testing; Tissues; United States National Institutes of Health; Universities; Wood material; Work; base; brain tissue; cellular imaging; clinical Diagnosis; drug development; experience; imaging modality; imaging system; infrared spectroscopy; instrument; instrumentation; irradiation; microscopic imaging; nano; nanoscale; particle; performance tests; product development; public health relevance; single cell analysis; small molecule; tool; user-friendly

 DESCRIPTION (provided by applicant): The goal of the proposed Phase II STTR research is to develop an imaging system capable of detecting and identifying biomolecules in cells and tissue sections with sub-cellular spatial resolution. The system combines an atomic force microscope (AFM) with mass spectrometry (MS) analysis and utilizes a unique tip-enhanced laser ablation (TELA) method developed in the Phase I portion of the research. The current technology limitation of imaging mass spectrometry is the fact that it is possible to obtain high-resolution images of small molecules and low-resolution images of large molecules, but it is not possible to both at once. The proposed instrument directly addresses this technology limitation. The TELA effect developed under Phase I as an off - line MS method will be developed under Phase II into an on-line instrument that integrates AFM and mass spectrometry imaging for both excellent spatial resolution and biomolecule mass spectrometry. The aims of the project are divided into 1) further refinement of TELA and development of the on-line TELA-MS interface, 2) integration of TELA and MS imaging, and 3) application to brain imaging. Refinement of the tip - enhanced laser ablation involves a study of the fundamental interaction between the pulsed ablation laser and the atomic force microscope tip. The optimized ablation system will be combined with an ultra-low flow electrospray ionization system in which charged particles from the electrospray source interact with the ablation plume to form biomolecule ions. Integration of the AFM and MS imaging systems will use four imaging modalities: spot-by-spot sampling, area sampling, area imaging, and depth profiling. The result is an AFM capable of both physical and chemical imaging of cells and tissue. The AFM TELA-MS device will be applied to detection of biomolecules in rat brain tissue. Spatial profiling of lipids, peptides, cocaine, and cocaine metabolites will be performed. The AFM TELA-MS system will be constructed by Anasys, based on past product development and manufacturing experience with nanoscale imaging systems such as atomic force microscopy and infrared spectroscopy chemical imaging using tunable pulsed laser sources. Mass spectrometry performance tests will be carried out on in the laboratory of Prof. Kermit Murray at Louisiana State University who is an expert in the development of laser ablation methods for the detection of biomolecules by mass spectrometry.

 描述（由适用提供）：拟议的II期STTR研究的目标是开发一个能够检测和鉴定具有亚细胞空间分辨率的细胞和组织切片中的生物分子的成像系统。该系统将原子力显微镜（AFM）与质谱分析（MS）分析相结合，并利用了在研究的I期部分中开发的独特的尖端增强激光消融方法（TELA）方法。成像质谱的当前技术局限性是，可以获得小分子和大分子的低分辨率图像的高分辨率图像的事实，但不可能一次同时获得。提议的仪器直接解决了这一技术限制。在I阶段作为OFF -LINE MS方法下开发的TELA效应将在II期下开发成一个在线仪器，该仪器将AFM和质谱成像整合为出色的空间分辨率和生物分子质谱法。该项目的目的分为1）TELA的进一步完善和在线TELA-MS接口的开发，2）TELA和MS成像集成，以及3）应用大脑成像。尖端增强激光消融的细化涉及研究脉冲消融激光与原子力显微镜尖端之间的基本相互作用。优化的消融系统将与超低流电喷雾电离系统结合使用，其中电喷雾源带电的颗粒与消融羽相互作用以形成生物分子离子。 AFM和MS成像系统的集成将采用四种成像方式：逐点采样，面积采样，区域成像和深度分析。结果是能够对细胞和组织进行物理和化学成像的AFM。 AFM Tela-MS设备将用于检测大鼠脑组织中的生物分子。将进行脂质，肽，可卡因和可卡因代谢物的空间分析。 AFM Tela-MS系统将由Anasys建立，该系统基于使用纳米级成像系统（例如原子力显微镜和红外光谱化学成像）的过去产品开发和制造经验，并使用可调脉冲激光源进行化学成像。路易斯安那州立大学Kermit Murray教授的实验室将进行质谱绩效测试，他是通过质谱法检测生物分子的激光消融方法的专家。