Eye Organ Specific Project

项目总结--眼器官专案。全面的三维生物分子分析 在组织内，需要协调来自许多分析技术的数据。我们将开发多式联运 直接从人类整个眼睛组织中获取空间特定的组学信息的策略。这项工作将 利用质谱学研究中心的独特仪器能力和专业知识 (Msrc)，其中斯普拉金斯、卡普里奥利、谢伊、古铁雷斯和范德普拉斯教授是关键人员。建房 在开发人体肾脏组织图谱的基础上，我们将把超高速、高空间 分辨率(&lt；10μm)MALDI时间高质量分辨率(&gt；40,000分辨率，&lt；2ppm质量的TOF IMS 准确性)，用于人眼的分子成像，具有前所未有的空间保真度和 分子特异性。MALDI timsTOF IMS的高通量能力将是产生高吞吐量的关键 来自统计相关数量的正常胰腺样本的空间分辨率三维分子图像。MALDI IMS将提供将离子图像链接到基于正交LC的离子图像所需的特异性和质量准确性 用于分子鉴定的碎裂实验。将使用空间特定的方式执行分段 以与IMS兼容的方式从组织中产生LC-MS/MS数据的表面采样方法 并且能够整合到HuBMAP组织图谱中。要最大化从成像中收集的信息 实验并为数据驱动的图像融合和数据中概述的三维重建奠定了基础 分析核心研究策略，我们还将开发收集各种基于显微镜的图像的方法 模式(如自体荧光和染色显微镜)和胰腺特异性，高度多元化 使用CODEX的免疫荧光。整个眼图将通过结合我们的多模式构建 使用体外3-D光学相干断层扫描(OCT)成像的IMS/显微镜管道。 通过使用我们的多模式管道，我们建议绘制眼睛细胞类型的空间关系图 从众多人口统计数据中的健康组织中提取眼睛的所有区域。目标1：创造一只眼睛- 具体的生物菌种收集和管理计划。目标2：建立一个分析前管道，用于 标准化样品制备，确定组织正常度，并提供质量控制指标。目标3： 开发一种用于眼部特异性3D分子成像的多模式表征流水线。目标4：扩展和 标准化眼睛特有的3-D分子成像。为了实现这些目标，我们召集了一个高度 互动式和成熟的研究团队，由眼科专业知识互补组成， 分析化学和数据科学。

PROJECT SUMMARY – Eye Organ Specific Project. Comprehensive 3-dimensional biomolecular analysis within tissues requires the coordination of data from many analytical technologies. We will develop a multimodal strategy to obtain spatially specific ‘omics’ information directly from human whole eye tissue. This work will leverage the unique instrumentation capabilities and expertise of the Mass Spectrometry Research Center (MSRC) in which Professors Spraggins, Caprioli, Schey, Gutierrez and Van de Plas are key personnel. Building from previous work developing atlases of human kidney tissue, we will integrate ultra-high speed, high spatial resolution (<10μm) MALDI timsTOF IMS with high mass resolution (>40,000 Resolving Power, <2 ppm mass accuracy) for molecular imaging of human eyes with an unprecedented combination of spatial fidelity and molecular specificity. The high-throughput capabilities of MALDI timsTOF IMS will be critical for generating high- spatial resolution 3-D molecular images from statistically relevant numbers of normal pancreas samples. MALDI IMS will provide the specificity and mass accuracy necessary to link ion images to orthogonal LC-based fragmentation experiments for molecular identification. Fragmentation will be performed using spatially specific surface sampling approaches to produce LC-MS/MS data from tissues in a manner that is compatible with IMS and capable of being integrated into HuBMAP tissue atlases. To maximize information gleaned from imaging experiments and lay the groundwork for data-driven image fusion and 3-D reconstruction outlined in the Data Analysis Core Research Strategy, we will also develop methods for collecting various microscopy-based image modalities (e.g. autofluorescence and stained microscopy) and pancreas specific, highly multiplexed immunofluorescence using CODEX. Whole eye atlases will be constructed by combining our multimodal IMS/Microscopy pipelines with ex vivo 3-D optical coherence tomography (OCT) imaging. Through the use of our multimodal pipeline, we propose to map spatial relationships of eye cell types from all regions of the eye within healthy tissue across a multitude of demographics. Aim 1: To create an eye- specific biospecimen collection and management plan. Aim 2: To establish a pre-analytical pipeline for standardizing sample preparation, determining tissue normalcy, and providing quality control metrics. Aim 3: To develop a multimodal characterization pipeline for eye-specific 3-D molecular imaging. Aim 4: To scale and standardize eye-specific 3-D molecular imaging. To accomplish these aims, we have assembled a highly interactive and established team of investigators consisting of complementary expertise in ophthalmology, analytical chemistry, and data science.