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（由申请人证明）：支撑II期Strer研究的目的是开发一个能够驱动和识别细胞中的Iomolecules的成像系统，该细胞与亚细胞空间空间空间分辨率IND tithe切片相结合。 AFM）具有质谱分析（MS）分析并利用在研究部分中开发的独特的尖端增强激光消融（TELA）方法。当前的成像质谱技术限制是可以获得高分辨率的质谱larges larges larges larges的图像立即限制了技术限制。质谱成像既有异常的空间分辨率和生物分子质量ctrometry ctrometry。 3）在脉冲消融激光器和原子力显微镜尖端之间的相互作用。对AFM和MS成像系统的融合将使用四个成像模态：逐个点采样，面积采样，面积成像和深度分析在大鼠脑组织中检测生物分子。使用脉冲激光源的化学成像质谱性能测试将通过质量特殊质量进行激光消融方法进行。