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

 描述(申请人提供)：拟议的第二阶段STTR研究的目标是开发一种能够以亚细胞空间分辨率检测和识别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)相结合，并利用在研究第一阶段开发的独特的尖端增强型激光消融(TELA)方法。目前成像质谱学的技术局限在于，可以获得小分子的高分辨率图像和大分子的低分辨率图像，但不可能同时获得两者。建议的仪器直接解决了这一技术限制。在第一阶段开发的Tela效应作为一种离线MS方法，将在第二阶段开发为一种在线仪器，它集成了原子力显微镜和质谱学成像，以获得卓越的空间分辨率和生物分子质谱学。这项计划的目标包括：1)进一步改善Tela和开发Tela-MS联机界面；2)整合Tela和MS成像；以及3)将其应用于脑成像。针尖增强激光烧蚀的改进包括研究脉冲烧蚀激光与原子力显微镜针尖之间的基本相互作用。优化的消融系统将与超低流量电喷雾电离系统相结合，在该系统中，来自电喷雾源的带电粒子与消融羽流相互作用，形成生物分子离子。AFM和MS成像系统的集成将使用四种成像方式：逐点采样、区域采样、区域成像和深度剖析。其结果是一种能够对细胞和组织进行物理和化学成像的原子力显微镜。AFM Tela-MS设备将用于检测大鼠脑组织中的生物分子。将对脂类、多肽、可卡因和可卡因代谢物进行空间分析。AFM Tela-MS系统将由Anasys公司基于过去在原子力显微镜和使用可调谐脉冲激光光源的红外光谱化学成像等纳米级成像系统的产品开发和制造经验构建。质谱学性能测试将在路易斯安那州立大学的Kermit Murray教授的实验室进行，他是开发激光消融方法的专家，用于用质谱学检测生物分子。

 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.