Bio-Mass Spec Microanalysis augments Optical Histology

生物质谱微量分析增强光学组织学

• 批准号: 7926642
• 负责人: Carey David Balaban
• 金额: $ 299.66万
• 依托单位: IONWERKS, INC.
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7926642
• 关键词: Ablation; Acids; Actins; Alloys; Basic Science; Biochemical; Biopsy Specimen; Caliber; Cations; Cell Volumes; Cell surface; Cells; Cellular Structures; Characteristics; Clinical Pathology; Coupled; DNA; Deposition; Development; Family; Film; Fullerenes; Goals; Growth; Height; Histology; Image; Implant; Ions; Lasers; Light; Lipids; Mammalian Cell; Mass Spectrum Analysis; Measurement; Metals; Methodology; Methods; Microscope; Microscopic; Molecular; Molecular Analysis; Molecular Profiling; Molecular Structure; Morphology; Optics; Particle Size; Peptides; Phenotype; Proteins; RNA; Sales; Science; Signal Transduction; Solid; Solutions; Specimen; Staining method; Stains; Structure of molecular layer of cerebellar cortex; Surface; Techniques; Technology; Testing; Time; Tissues; Variant; Width; Work; analytical method; base; design; implantation; infancy; innovation; instrument; ion mobility; ionization; molecular phenotype; monolayer; nanoparticulate; novel strategies; organic acid; particle; public health relevance; rapid technique; submicron

DESCRIPTION (provided by applicant): Meaningful molecular analysis of single mammalian cells by submicron focused laser desorption mass spectrometry has been an elusive goal. However, such microanalysis has been achieved with our unique MALDI-Ion Mobility-orthogonal Time of Flight Mass Spectrometry (MALDI-IM-oTOFMS). Applying submonolayers of special nanoparticulate matrices (NP) onto sub cellular tissue volumes enables a micro focused laser to desorb neutral analyte molecules (which are then post-ionized with a second laser and mass analyzed). Thus, the applicants' main goal now is the creation and optimization of new nanoparticulate matrices and techniques for rapid dissemination of NPs into sub cellular volumes. We will (1) demonstrate sub cellular molecular profiling by NP implantation and compare against matrices used in MALDI tissue imaging (2) design nanoparticulates which promote the non-destructive laser desorption of neutral analytes which can then be post-ionized with a second laser, and (3) demonstrate molecular depth profiling of cells through repeated cycles of matrix implantation followed by molecular analysis by MALDI-IM-oTOFMS (with post- ionization). This approach will be compared with several standard histological stains for DNA, RNA and f-actin (among others). We consider these developments to be critical bridging technologies for transforming quantitative micro-analytical methods into biochemical imaging of sub cellular molecular structure useful for basic research and pathological analysis. PUBLIC HEALTH RELEVANCE: The Ionwerks nanoparticulate implanter is now offered for sale, and the innovations described by the applicants can be rapidly tested and integrated to augment the power of an already useful instrument and technique which is in its commercial infancy. The rapid implantation of nanoparticulate matrices is essential technology for intracellular analysis and intracellular analysis is the basis for molecular phenotyping. Thus achieving cellular level mass spectrometry-based molecular phenotyping seems feasible and would be an important development in biomedical sciences and clinical pathology. Simple approaches, such as overlays of the mass spec image and light (or confocal) microscopic images should provide fast, streamlined, broad molecular phenotyping of even small or limited biopsy samples.

描述（由申请人提供）：通过亚微米聚焦激光解吸质谱法对单个哺乳动物细胞进行有意义的分子分析一直是一个难以实现的目标。然而，这种微量分析已经实现了我们独特的MALDI-离子迁移率-正交飞行时间质谱（MALDI-IM-oTOFMS）。将特殊纳米颗粒基质（NP）的亚单层应用到亚细胞组织体积上，使微聚焦激光能够解吸中性分析物分子（然后用第二激光进行后电离并进行质量分析）。因此，申请人现在的主要目标是创造和优化新的纳米颗粒基质和技术，用于将NP快速散布到亚细胞体积中。我们将（1）通过NP植入证明亚细胞分子谱，并与MALDI组织成像中使用的基质进行比较（2）设计促进中性分析物的非破坏性激光解吸的纳米颗粒，然后可以用第二激光后电离中性分析物，和（3）通过基质植入的重复循环，然后通过MALDI-IM-oTOFMS进行分子分析，证明细胞的分子深度分布（具有后电离）。该方法将与用于DNA、RNA和f-肌动蛋白（以及其他）的几种标准组织学染色剂进行比较。我们认为这些发展是关键的桥梁技术，将定量微量分析方法转化为对基础研究和病理分析有用的亚细胞分子结构的生物化学成像。 公共卫生相关性：Ionwerks纳米颗粒过滤器现在可供销售，并且申请人所描述的创新可被快速测试和集成，以增强已经有用的仪器和技术的能力，该仪器和技术处于商业婴儿期。纳米颗粒基质的快速植入是细胞内分析的基本技术，而细胞内分析是分子表型分析的基础。因此，实现基于细胞水平质谱的分子表型分析似乎是可行的，并且将是生物医学科学和临床病理学的重要发展。简单的方法，如质谱图像和光（或共聚焦）显微镜图像的叠加，应该提供快速，精简，广泛的分子表型，即使是小的或有限的活检样本。

项目成果

Laser Desorption/Ionization Mass Spectrometric Imaging of Endogenous Lipids from Rat Brain Tissue Implanted with Silver Nanoparticles.

• DOI: 10.1007/s13361-017-1665-4
• 发表时间: 2017-08
• 期刊: Journal of the American Society for Mass Spectrometry
• 影响因子: 3.2
• 作者: Muller L;Baldwin K;Barbacci DC;Jackson SN;Roux A;Balaban CD;Brinson BE;McCully MI;Lewis EK;Schultz JA;Woods AS
• 通讯作者: Woods AS

Imaging of lipids in rat heart by MALDI-MS with silver nanoparticles.

• DOI: 10.1007/s00216-013-7525-6
• 发表时间: 2014-03
• 期刊: Analytical and bioanalytical chemistry
• 影响因子: 4.3
• 作者: Jackson SN;Baldwin K;Muller L;Womack VM;Schultz JA;Balaban C;Woods AS
• 通讯作者: Woods AS

Chronic ethanol consumption profoundly alters regional brain ceramide and sphingomyelin content in rodents.

• DOI: 10.1021/cn500174c
• 发表时间: 2015-02-18
• 期刊: ACS CHEMICAL NEUROSCIENCE
• 影响因子: 5
• 作者: Roux, Aurelie;Muller, Ludovic;Jackson, Shelley N.;Baldwin, Katherine;Womack, Virginia;Pagiazitis, John G.;O'Rourke, Joseph R.;Thanos, Panayotis K.;Balaban, Carey;Schultz, J. Albert;Volkow, Nora D.;Woods, Amina S.
• 通讯作者: Woods, Amina S.

MALDI/post ionization-ion mobility mass spectrometry of noncovalent complexes of dopamine receptors' epitopes.

多巴胺受体表位的非共价复合物的MALDI/电离后离子迁移率质谱。

• DOI: 10.1021/pr301004w
• 发表时间: 2013-04-05
• 期刊: Journal of proteome research
• 影响因子: 4.4
• 作者: Woods AS;Jackson SN;Lewis EK;Egan T;Muller L;Tabet JC;Schultz JA
• 通讯作者: Schultz JA