3-Dimensional Imaging of Biomaterials on the Nanoscale with Mass Spectrometry

利用质谱法对纳米级生物材料进行 3 维成像

• 批准号: 7827728
• 负责人: NICHOLAS WINOGRAD
• 金额: $ 161.34万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-13 至 2012-05-12
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7827728
• 关键词: 3-Dimensional; Air; Anabolism; Asthma; Biocompatible Materials; Biological; Biological Assay; Biomaterials Research; Brain; Cells; Chemicals; Communities; Complex; Consumption; Country; Defect; Development; Dimensions; Disease; Environmental Impact; Financial Support; Freezing; Functional disorder; Funding; Heart; Hela Cells; Hypersensitivity; Image; Individual; Ions; Laboratories; Lateral; Learning; Lipid Biochemistry; Lipids; Liposomes; Mass Spectrum Analysis; Memory; Methods; Neurotransmitters; Pathway interactions; Pharmaceutical Preparations; Purines; Research; Research Personnel; Resolution; Sampling; Science; Shipping; Ships; Stroke; Techniques; Time; Tissues; United States National Institutes of Health; Universities; Vacuum Pumps; Vesicle; base; bioimaging; cellular imaging; chemotherapeutic agent; cryogenics; design; instrument; interest; mass analyzer; mass spectrometer; membrane model; nanoscale; neurotransmission; novel; public health relevance; purine; square foot; tool

DESCRIPTION (provided by applicant): This proposal describes a request for funds to purchase a newly developed secondary ion mass spectrometer for 3-dimensional chemical imaging of biological cells and tissue. The instrument is based upon the use of a micro-focused ion beam consisting of energetic C60+ or Au3+ cluster ions to desorb biomolecules from a well-defined x,y coordinate into a novel TOF-TOF mass analyzer. Information about the 3rd dimension is obtained by erosion of the sample under high fluence ion bombardment conditions. This instrument is essential for the growing imaging mass spectrometry community because of its sophisticated cryogenic sample handling, greatly increased sensitivity and ms/ms capability. In addition, because of the unique design features, the instrument provides unprecedented lateral resolution of 300 nm for C60, and 50 nm for Au3, allowing sub-cellular imaging. These features will accelerate the research objectives of many NIH supported investigators interested in the cellular localization of drugs, metabolites and lipids. Representative projects include elucidation of de novo purine biosynthetic pathways in HeLa cells, assay of single vesicles in neurotransmitters, lipid biochemistry at model membranes, heart tissue and in liposomes. Five different collaborators from three Universities are involved in this effort, a situation made possible by the unique cryogenic sample handling associated with the mass spectrometer that allows frozen samples to be shipped across the country by air express. The instrument will initially reside in the PI's laboratory and be guided by an Oversight Committee. Financial support will come from the PI and from contributions from the collaborators. After the initial period of adjustment, the instrument will transition to an emerging Millennium Science Complex, a 265,000 square foot facility dedicated to biomaterials research. Although the initial number of collaborators is not large due to the complexity of each individual project and time limitations, it is anticipated that the results of this research will stimulate other groups to acquire this capability, bringing the technique to a wide swath of NIH investigators. PUBLIC HEALTH RELEVANCE: The 3-Dimensional Chemical Imager is a unique nanoscale tool for bioimaging that will provide basic information about mechanisms of neurotransmission, purine biosynthesis and lipid biochemistry. Results of these studies are proposed to impact a variety of disease states including learning memory and brain dysfunction, stroke, aortal valve defects, asthma and allergies, and the development of chemotherapeutic agents. The results will be overseen by a highly skilled technical staff that will be expanded over time to accommodate new users and new scientific endeavors. The environmental impact of mass spectrometry is relatively neutral with respect to most wet chemical or other instrumental methods. Samples are analyzed without significant chemical usage, and power consumption, associated mostly with vacuum pumps, is modest during and after spectrometer evacuation.

描述（由申请人提供）：该提案描述了一项要求购买新开发的二级离子质谱仪的资金请求，用于生物细胞和组织的三维化学成像。该仪器基于使用的微焦离子束的使用，该离子束由能量的C60+或AU3+簇离子组成，以从定义明确的X，y坐标为新型TOF-TOF-TOF质量分析仪中取消生物分子。有关第三维的信息是通过在高频离子轰击条件下侵蚀样品获得的。由于其复杂的低温样品处理，灵敏度大大提高和MS/MS功能，因此该仪器对于不断增长的成像质谱群落至关重要。此外，由于具有独特的设计功能，该仪器为C60提供了前所未有的侧向分辨率，AU3提供了50 nm，允许亚细胞成像。这些功能将加速许多NIH支持对药物，代谢产物和脂质细胞定位感兴趣的研究者的研究目标。代表性的项目包括阐明新嘌呤生物合成途径在HeLa细胞中，神经递质中单囊泡的测定，模型膜，心脏组织和脂质体的脂质生物化学。来自三所大学的五个不同合作者参与了这项工作，这种情况通过与质谱仪相关的独特低温样本处理使这种情况成为可能，这使得允许Air Express在全国范围内运送冷冻样本。该工具最初将居住在PI的实验室中，并由监督委员会指导。财政支持将来自PI和合作者的贡献。调整初期后，该仪器将过渡到新兴的千年科学综合体，这是一个专门用于生物材料研究的265,000平方英尺的设施。尽管由于每个项目和时间限制的复杂性，合作者的初始数量并不大，但预计这项研究的结果将刺激其他群体获得该能力，从而将技术带入广泛的NIH研究人员。 公共卫生相关性：三维化学成像仪是一种独特的纳米级工具，可提供有关神经传递，嘌呤生物合成和脂质生物化学机制的基本信息。提出这些研究的结果会影响各种疾病状态，包括学习记忆和脑功能障碍，中风，主动脉瓣缺陷，哮喘和过敏以及化学治疗剂的发展。结果将由一位高技能的技术人员监督，随着时间的流逝，该公司将扩大，以适应新的用户和新的科学努力。相对于大多数湿化学或其他仪器方法，质谱的环境影响相对中立。在没有明显的化学使用情况下分析样品，并且在光谱仪撤离期间和之后，主要与真空泵相关的功耗是适度的。