Spatial Analysis of Molecules in Tissue using MALDI-MS

描述（由申请人提供）：拟议研究的目标是开发高分辨率3-D成像质谱（IMS）技术，使其能够在实际时间框架内提供高分辨率图像。高空间分辨率IMS （~10 μ m）将与提供1 μ m矩阵晶体尺寸和高速激光器的组织制备方案一起使用。3khz)，以有效和及时的方式提供多个串行段的数据。计划开发的方案将允许多肽、蛋白质、脂质、药物和其他小分子在组织体积中以3d方式进行跟踪。此外，工作计划完成重要的生物计算工作，以处理数据维数和尺寸减小，离子强度的标准化，用于在单个二维实验中相对定量的图像，以及用于三维重建的组织体积的连续切片，具有解剖保真度的切片注册，以及在实际时域中生成单个分子的三维图像。这项技术将解决几个重要的生物学问题；1)评估小鼠大脑纹状体中纹状体的3-D分子分布和边界，与神经精神疾病（如帕金森病、亨廷顿氏病和强迫症）有关的亚结构；2)在肿瘤和微环境中，在小鼠视神经模型中，与脑图谱中现有的3-D显微镜图像相关联，对胶质瘤生长、进展和维持过程中的分子事件进行3-D映射；3)三维IMS数据与其他成像方式的协同配准；PET成像造影剂的配置和代谢研究，MRI扩散加权成像中分子方面的阐明，以及IMS与MR弛豫数据获得的分子事件的共配准/相关性。这项工作将汇集几种强大的成像模式，并通过每种模式的独特属性，在理解整个组织动力学方面获得组合优势。其中一个目标是，计划将显微镜和免疫组化测量、核磁共振和IMS结合起来，作为一个单一的3-D体积的一部分，可以使用标准的互联网工具轻松地进行查询。

DESCRIPTION (provided by applicant): The goal of the proposed studies is to develop high resolution 3-D Imaging Mass Spectrometry (IMS) technology to a level that it is capable of delivering high resolution images in a practical time frame. High spatial resolution IMS (~10 um) will be employed with tissue preparation protocols that provide 1 um matrix crystal sizes and high speed lasers (.3 kHz) to provide data on multiple serial sections in a efficacious and timely manner. It is planned to develop the protocols that would allow peptides, proteins, lipids and drugs and other small molecules to be tracked in a 3-D manner in a tissue volume. In addition, work is planned to accomplish the significant biocomputational effort needed to deal with data dimensionality and size reduction, normalization of ion intensities for the relative quantitation of images both within a single 2-D experiment and across serial sections of a tissue volume for 3-D reconstruction, registration of the sections with anatomic fidelity, and generation of a 3-D images of individual molecules in a practical time domain. Several important biological problems will be addressed with this technology; 1) assessment of the 3-D molecular distributions and boundaries of striosomes in the striatum of the mouse brain, substructures implicated in neuropsychiatric disorders such as Parkinsonism, Huntington's Disease, and obsessive-compulsive disorder, 2) 3-D mapping of molecular events in glioma growth, progression, and maintenance, both in the tumor and microenvironment, in the optic nerve of a mouse model with linkage to existing 3-D microscopy images in the brain atlas, 3) Co-registration of 3-D IMS data with other imaging modalities; studies of the disposition and metabolism of contrast agents used for PET imaging, the elucidation of molecular aspects in diffusion weighted imaging by MRI, and the co-registration/correlation of molecular events obtained by IMS with MR relaxation data. This work will bring together several powerful imaging modalities and, through the unique attributes of each, gain a combinatorial advantage in understanding whole tissue dynamics. In one aim, it is planned to combine microscopy and IHC measurements, MRI and IMS as part of a single 3-D volume that is easily interrogated using standard internet tools.