RVC - Polymer Plate for improved biomedical analyses by tissue imaging MALDI-TOF

描述：本提案的目的是尝试用基质辅助激光解吸电离成像质谱法（MALDI-IMS）分析生物组织的新模式。MALDI-IMS是一种强大的分析技术，能够产生~ 50-100微米分辨率的图像，可以直接从生物组织切片中按质量过滤。该技术的应用范围包括对组织中生物分子和药物的分布进行分类，发现生物标志物，肿瘤分类，阐明发病机制，研究发育生物学等。该建议概述了一种旨在从几个方面改进MALDI-IMS分析的新技术。该技术的基础是新型底物的构建，旨在选择性地从组织中提取、捕获和富集感兴趣的分析物，同时保持样品的空间分辨率，其格式将直接与质谱仪接口。本文概述的方法直接解决了当前MALDI-IMS分析中最公认和最一致的弱点，并将通过为现有应用程序提供更多细节、平衡和多功能性以及提供令人兴奋的扩展潜力来增强MALDI-IMS性能。该项目初始阶段的具体目标是基材建设和实验程序。每个目标都对产品可行性证明至关重要，它们允许与当前最先进的MALDI-IMS技术进行量化评估和比较性能。这里假设，所提出的技术将提高MALDI-IMS的检测阈值，通过实现对目标分析物的选择性，同时去除干扰。它将通过允许更合适的分析物/基质共结晶条件来提高灵敏度，并通过为质谱分析提供更理想的环境来提高数据质量。它将通过呈现一个基底来稳定组织图像，该基底将准确地锁定组织的空间完整性，其分辨率等于或大于当前协议所能达到的分辨率。它将通过分离分析萃取和基质沉积的步骤来增加样品内的一致性和样品间的可重复性，并为完成这些任务提供更系统和优化的方法。它将通过允许样本操作而不干扰空间完整性来扩大实验能力。它将通过保存母样品来扩大实验能力，允许对同一样品进行多次，重复或不同的分析。这将扩大实验能力，分析各种形状和形式的组织，甚至可以分析活体组织。它将通过消除对低温切片组织标本的限制来简化实验方案，并有望通过消除对专业和昂贵设备的需求来扩大对MALDI-IMS技术的参与。原型衬底由Duocel(R)网状玻璃碳（RVC）泡沫与多孔聚合物单体结合而成。用于开发这些独特基板的材料是市售的。在板结构中使用的化学物质是很容易理解的，所需的反应物是市售的。将被尝试的组织样本操作记录在文献中。这里展示的初步数据令人鼓舞。

DESCRIPTION: The objective of this proposal is to attempt a new mode of analysis for biological tissue by matrix-assisted laser desorption ionization imaging mass spectrometry (MALDI-IMS). MALDI-IMS is a powerful analytical technique capable of generating ~ 50-100 micrometer resolution images that can be filtered by mass, directly from biological tissue sections. Applications for the technique range from cataloging the distribution of bio-molecules and pharmaceuticals in tissues, to biomarker discovery, tumor classification, elucidation of pathogenesis, studies of developmental biology, etc. This proposal outlines a new technology aimed at improving MALDI-IMS analyses on several fronts. The basis of the technology is the construction of novel substrates designed to selectively extract, capture and enrich analyte(s) of interest from tissue, while preserving sample spatial resolution, in a format that will interface directly with the mass spectrometer. The methodology outlined here-in directly addresses the most recognized and consistent points of weakness in current MALDI-IMS analyses and will serve to enhance MALDI-IMS performance by providing more detail, balance and versatility to existing applications as well as providing exciting potential for expansion. The specific aims for this initial phase of the project target substrate construction and experimental procedure. Each aim is critical to proof- of-product feasibility and they allow for quantifiable evaluation and comparative performance with current, state-of-the-art MALDI-IMS technology. It is hypothesized here that the proposed technology will increase the detection thresholds of MALDI-IMS by enabling selectivity for targeted analyte(s) with simultaneous removal of interferents. It will increase sensitivity by allowing more appropriate conditions for analyte / matrix co-crystallization and it will increase data quality by providing an environment more optimal for mass spectrometry. It will stabilize the tissue image by presenting a substrate that will accurately lock-in the spatial integrity of the tissue at a resolution equal-to or greater- thn what is achievable by current protocols. It will increase intra-sample consistency and inter-sample reproducibility by decoupling the steps of analyst extraction and matrix deposition and providing a more systematic and optimized means for accomplishing these tasks. It will expand experimental capability by allowing sample manipulation without disturbing the spatial integrity. It will expand experimental capability by preserving the parent sample allowing for multiple, repetitive or divergent analyses to be performed on the same specimen. It will expand experimental capability by enabling the analysis of tissues of diverse shapes and forms and even allow for the analysis of living tissue. It will simplify experimental protocol by removing restrictions for cryogenically sliced tissue specimens and it will hopefully serve to expand participation in MALDI-IMS technology by lifting the need for specialized and expensive equipment. The prototype substrates are composed of a Duocel(R) reticulated vitreous carbon (RVC) foam combined with a porous polymer monolith. The materials used to develop these unique substrates are commercially available. The chemistry to be employed in plate construction is well understood and the reactants needed are commercially available. The tissue sample manipulations that will be attempted are documented in the literature. The preliminary data demonstrated here-in are encouraging.