Belizário José E, Sangiuliano Beatriz A, Perez-Sosa Marcela, Santos Beatriz V, Machado-Santelli Gláucia
Department of Pharmacology, Institute of Biomedical Sciences, University of Sao Paulo, São Paulo, SP, Brazil.
Department of Cell Biology and Development, Institute of Biomedical Sciences, University of Sao Paulo, São Paulo, SP, Brazil.
Discov Med. 2015 Dec;20(112):393-401.
Many molecular mechanisms in complex biological processes of diseases cannot be fully understood without direct visualization. In the last decades, advances in imaging principles and technologies have expanded our ability to capture and analyze the morphology of tissues and cells' components on conventional widefield optical and fluorescence microscopies and their derivatives confocal and multiphoton fluorescence laser-scanning microscopes, as well as transmission electron microscopes. Innovative imaging technologies are now emerging for constructing fine structural features, precise localization and the dynamic interplay of single and macromolecular assemblies that drive cell growth, differentiation and cell death as well as stromal and chromatin remodeling within many cellular context. The newer super-resolution microscopies capture images with unprecedented sensitivity and clarity allowing the exploration of interactions between individual molecules with a distance resolution as low as 20 nm. But these techniques are not robust enough to quantitate molecules on a genome-wide scale. Mass spectrometry imaging is a high-throughput chemical imaging technique for the identification, quantitation and distribution of proteins, lipids and chemical metabolites at picomol level within a single-cell and complex multicellular tissue. Here we provide an overview on imaging instrumentations and computational platforms to store, data mining, analyze and retrieving genomic, proteomic and immunohistochemistry digital image information which are available for multilevel academic-private collaborations. The expansion of these data sets will lead to a merge picture from it we will retrieve knowledge for more rational-design systems to basic and clinical research in near future.
在疾病复杂的生物学过程中，许多分子机制若没有直接可视化就无法被完全理解。在过去几十年里，成像原理和技术的进步扩展了我们在传统宽视野光学显微镜、荧光显微镜及其衍生的共聚焦和多光子荧光激光扫描显微镜以及透射电子显微镜下捕捉和分析组织形态及细胞成分的能力。如今，创新成像技术正在兴起，用于构建精细的结构特征、精确的定位以及单个和大分子组装体的动态相互作用，这些组装体驱动细胞生长、分化和细胞死亡，以及许多细胞环境中的基质和染色质重塑。更新的超分辨率显微镜以前所未有的灵敏度和清晰度捕捉图像，使得能够探索距离分辨率低至20纳米的单个分子之间的相互作用。但这些技术在全基因组范围内对分子进行定量分析的能力还不够强大。质谱成像技术是一种高通量化学成像技术，用于在单细胞和复杂多细胞组织中以皮摩尔水平鉴定、定量和分布蛋白质、脂质和化学代谢物。在此，我们概述了成像仪器和计算平台，用于存储、数据挖掘、分析和检索基因组、蛋白质组和免疫组织化学数字图像信息，这些信息可用于多层次的学术 - 私人合作。这些数据集的扩展将形成一幅融合的图像，从中我们将在不久的将来为基础和临床研究获取知识，以设计更合理的系统。
