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（Louisiana State University）和贝勒大学（Baylor University）获得了一项奖励，用于开发一种利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，低成本，易于使用，将为基础和转化生物学研究的广泛研究人员带来高性能的非靶向生化成像。该仪器将用于结合采样和成像工作流程，具有定位、识别和量化组织中的生物分子的能力。质谱法在体外成像方法中具有非靶向性的独特优势；因此，它具有检测脂质、代谢物、肽和蛋白质的能力，而无需事先了解其成分。该仪器解决了亚微米精度的大生物分子电离的关键技术障碍，这是化合物的细胞定位所必需的。受益于该仪器的用户群体包括需要空间定位的化合物鉴定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一种能够分析多种生化类的仪器。转化研究将受益于一种坚固、高灵敏度的仪器，相对较低的成本和易用性将使其进入研究中心，而这些研究中心通常没有这种能力。质谱成像项目将利用STEM教育资源进行高中、本科、研究生和博士后的教育和培训。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。参加路易斯安那州生物医学研究网络暑期研究项目的本科生研究人员，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机会。目前的质谱成像技术需要在二次离子质谱（SIMS）和基质辅助激光解吸电离（MALDI）之间进行折衷，次级离子质谱（SIMS）使用精确聚焦的离子束对小分子进行成像，而基质辅助激光解吸电离（MALDI）使用较少聚焦但更温和的激光束对大分子进行成像。提出的研究通过使用高度聚焦的深紫外烧蚀激光器与双激光MALDI （MALDI-2）的紫外后电离激光器相结合，消除了对空间分辨率和质量范围妥协的需要。该方法利用MALDI效应对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、可靠和相对低成本的质谱技术，有可能将仪器的功能带给更多的生物成像研究人员。MALDI-2成像与深紫外激光烧蚀取样相结合，用于液相色谱串联质谱分析。该项目的成果将通过各种社交媒体平台以及传统的出版物和会议演讲方法进行传播。将通过专门的网站和社交媒体建立在线存在，该网站和社交媒体将在仪器上发布照片、视频和更新。该网站将兼容移动设备，并提供仪器运行的照片和视频以及所获得的数据。该站点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一个原型深紫外烧蚀MALDI-2成像质谱仪，空间分辨率为1µm，质量范围大于10,000 Da。该仪器将能够快速成像，易于样品制备，易于数据采集，可与传统的MALDI成像质谱相媲美。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。