IIBR Instrumentation: Sub-micrometer Resolution Mass Spectrometry Imaging by Deep-UV Laser Ablation and Post-ionization

IIBR 仪器：通过深紫外激光烧蚀和后电离进行亚微米分辨率质谱成像

• 批准号: 1951447
• 负责人: Kermit Murray
• 金额: $ 83.67万
• 依托单位: Louisiana State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1951447&HistoricalAwards=false
• 关键词: IIBR; Instrumentation; Sub; micrometer; Resolution

An award is made to Louisiana State University and Baylor University to develop an instrument for imaging biomolecules in tissue that uses mass spectrometry and has both high spatial resolution and high mass range. The instrument is simple, low-cost, and easy to use and will bring high-performance untargeted biochemical imaging to a broad range of researchers in fundamental and translational biological research. This instrument will be used in a combined sampling and imaging workflow with the ability to localize, identify, and quantify biomolecules in tissue. Mass spectrometry has the unique advantage among in vitro imaging methods in that it is un-targeted; thus, it has the capability of detecting lipids, metabolites, peptides and proteins without prior information about the components. This instrument addresses a critical technological barrier of large biomolecule ionization at submicrometer precision that is necessary for cellular localization of compounds. The user community benefiting from the proposed instrument includes researchers in fundamental and applied areas that require spatially localized chemical compound identification and quantification. Basic biological research will benefit from an instrument that can analyze a wide range of biochemical classes. Translational research will benefit from an instrument that is robust and highly sensitive and the relatively low cost and ease of use will bring it to research centers where this capability is not typically found. The mass spectrometry imaging project will leverage STEM education resources for high-school, undergraduate, graduate, and postdoctoral education and training. The project will provide research opportunities for students participating in the NSF S-STEM Scholars program and the Louis Stokes Louisiana Alliance for Minority Participation. Opportunities will be available for undergraduate researchers from the State participating in the Louisiana Biomedical Research Network summer research program and for local middle and high school science teachers through the LSU Cain Center for STEM Literacy.Current mass spectrometry imaging techniques require a compromise between secondary ion mass spectrometry (SIMS) that uses a precisely focused beam of ions to image small molecules and matrix-assisted laser desorption ionization (MALDI) that uses a less-focused but gentler laser beam to image large molecules. The proposed research eliminates the need for this spatial resolution and mass range compromise through the use of a highly focused deep-UV ablation laser combined with an ultraviolet post-ionization laser for two-laser MALDI (MALDI-2). The approach uses the MALDI effect on ablated tissue and matrix material and therefore has the mass range and gentle ionization properties of MALDI. Furthermore, it uses simple, robust, and relatively low-cost mass spectrometry technology that has the potential to bring the capabilities of the instrument to more biological imaging researchers. MALDI-2 imaging is combined with deep-UV laser ablation sampling for liquid chromatography tandem mass spectrometry analysis. The results of the project will be disseminated using a variety of social media platforms as well as traditional methods of publications and conference presentations An online presence will be established through a dedicated website and social media that will post photos, videos, and updates on the instrument. The website will be mobile device compatible and feature photos and videos of the instrument in operation as well as the data obtained. The site will also include instructions and protocols, example data, and instrument control and analysis software. The anticipated outcome of this project is a prototype deep-UV ablation MALDI-2 imaging mass spectrometer with 1 µm spatial resolution and greater than 10,000 Da mass range. The instrument will be capable of imaging with the speed, ease of sample preparation, and ease of data acquisition that is comparable to conventional MALDI imaging mass spectrometry.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

路易斯安那州立大学和贝勒大学获得了一项奖项，开发一种使用质谱法对组织中的生物分子进行成像的仪器，该仪器具有高空间分辨率和高质量范围。该仪器简单、成本低、易于使用，将为广大基础和转化生物学研究人员带来高性能的非靶向生化成像。该仪器将用于组合采样和成像工作流程，能够定位、识别和量化组织中的生物分子。质谱技术在体外成像方法中具有独特的优势，即非靶向性；因此，它能够检测脂质、代谢物、肽和蛋白质，而无需事先了解成分信息。该仪器解决了亚微米精度大生物分子电离的关键技术障碍，这是化合物细胞定位所必需的。受益于所提出的仪器的用户社区包括需要空间局部化合物识别和定量的基础和应用领域的研究人员。基础生物学研究将受益于能够分析多种生化类别的仪器。转化研究将受益于坚固且高度灵敏的仪器，并且相对较低的成本和易用性将使其带到通常不具备这种能力的研究中心。质谱成像项目将利用 STEM 教育资源进行高中、本科、研究生和博士后教育和培训。该项目将为参加 NSF S-STEM 学者计划和路易斯·斯托克斯路易斯安那州少数民族参与联盟的学生提供研究机会。来自该州参加路易斯安那生物医学研究网络夏季研究计划的本科研究人员以及通过路易斯安那州立大学凯恩 STEM 素养中心为当地初中和高中科学教师提供的机会。当前的质谱成像技术需要在二次离子质谱 (SIMS) 和基质辅助激光之间进行折衷，二次离子质谱成像技术使用精确聚焦的离子束对小分子进行成像 解吸电离 (MALDI) 使用聚焦较少但较温和的激光束对大分子进行成像。拟议的研究通过使用高度聚焦的深紫外烧蚀激光器与紫外后电离激光器相结合的双激光器 MALDI (MALDI-2)，消除了这种空间分辨率和质量范围折衷的需要。该方法利用 MALDI 效应作用于消融组织和基质材料，因此具有 MALDI 的质量范围和温和的电离特性。此外，它使用简单、稳健且成本相对较低的质谱技术，有可能为更多的生物成像研究人员带来该仪器的功能。 MALDI-2 成像与深紫外激光烧蚀采样相结合，用于液相色谱串联质谱分析。该项目的结果将通过各种社交媒体平台以及传统的出版物和会议演示方法进行传播。将通过专门的网站和社交媒体建立在线展示，发布仪器的照片、视频和更新。该网站将与移动设备兼容，并提供仪器运行中的照片和视频以及获得的数据。该网站还将包括说明和协议、示例数据以及仪器控制和分析软件。该项目的预期成果是原型深紫外烧蚀 MALDI-2 成像质谱仪，具有 1 µm 空间分辨率和大于 10,000 Da 质量范围。该仪器将能够以与传统 MALDI 成像质谱仪相当的速度、样品制备简便性和数据采集简便性进行成像。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。