Integrated multi-dimensional molecular organ imaging

集成多维分子器官成像

• 批准号: MR/K015710/1
• 负责人: Jeffrey Pollard
• 金额: $ 205.91万
• 依托单位: University of Edinburgh
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2013
• 资助国家: 英国
• 起止时间: 2013 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FK015710%2F1
• 关键词: Integrated; multi; dimensional; molecular; organ

The development of innovative optical imaging technologies has led to a revolution in light microscopy heralding the development of a generation of microscopes that promise powerful new applications for biomedical research. We believe our plans for refinement of commercially available instrumentation, development of novel probes and comprehensive testing in complex biological applications will ensure our advanced light microscopy 'toolkit' can be exploited to deliver innovative research and improved health outcomes.To maximise our contribution to the innovation and application of advanced imaging we focus on intra-vital imaging as this platform allows detailed investigation of biological events at sub-cellular resolution in living specimens at the greatest possible penetration depth. We will deploy a state-of-the-art intra-vital light microscope to acquire images of specifically labelled or label-free molecules up to one millimetre into tissue(s) of live animals typically mouse or zebrafish. Our innovative approach includes complementing these images with 3D mapping in order to provide a much more complete view of organ function in health and disease.Effective, informative, methods for labelling cells prior to intra-vital imaging remains a significant challenge in the wider application of these methods to non-transgenic models. In order to increase the reach of our intra-vital imaging methods we will employ and evaluate novel fluorescent 'Smartprobes'. These Smartprobes are already being used in Edinburgh to image key biological and pathological events in human lungs through cutting-edge fibre-optic microscopy, thus providing a direct translational pathway from planned studies in zebrafish and mice to human diseased organs in life (the technology is readily applicable to the digestive tract and the reproductive system). Our group has already established a strong track record in the use of a wide range of model animals that express fluorescently labelled molecules of interest in key target cells/organs. This expertise will guarantee we develop robust methodology focussed on scientifically challenging questions, such as: What is the involvement of macrophages in the development of breast cancer? How do neurons and glial cells interact in the development, function and repair of the nervous system? How do sex steroid hormones alter cell behaviour in the uterus and how can this be modulated? What is the distribution of central neuronal proteins in the brains of normal compared to model mice suffering from brain disorders?Excitement surrounding the application of Intra-vital imaging to biomedical research is based on its capacity to track cells and their contents within live tissues offering a novel platform for the study of dynamic changes in protein distribution, molecular interactions and tissue composition. Currently, such observations are only possible at single locations within a tissue - our goal is to expand data-acquisition to whole organs, e.g. to interrogate the distribution of synaptic proteins in a mouse brain. To achieve this, we will integrate our intra-vital techniques with systematic 3D image acquisition on a fast confocal microscope, capable of scanning whole sectioned organs. The resulting series of 3D-images will be archived to ensure data preservation and will be made available to the wider research community via searchable web-based organ libraries. These molecular 'organ maps' detailing the location of specific labelled proteins will allow us to refine subsequent 'focussed' functional intra-vital imaging and provide high-content 3D organ maps with functional measurements from live specimens.These innovative studies will provide data for our translational research pipeline and inform development of improved diagnostics and therapies for health problems including cancer, multiple sclerosis, infertility, cardiovascular and neurodegenerative diseases.

创新光学成像技术的发展引发了光学显微镜的革命，预示着新一代显微镜的发展，为生物医学研究带来了强大的新应用。我们相信，我们对商用仪器的完善、新型探针的开发以及复杂生物应用中的全面测试的计划将确保我们先进的光学显微镜“工具包”可用于提供创新研究和改善健康结果。为了最大限度地为先进成像的创新和应用做出贡献，我们专注于活体成像，因为该平台允许在活体样本中以亚细胞分辨率详细研究生物事件。 最大可能的穿透深度。我们将部署最先进的活体光学显微镜，以获取活体动物（通常是小鼠或斑马鱼）组织中长达一毫米的专门标记或无标记分子的图像。我们的创新方法包括用 3D 绘图补充这些图像，以便提供健康和疾病中器官功能的更完整视图。在活体成像之前标记细胞的有效、信息丰富的方法仍然是这些方法更广泛应用于非转基因模型的重大挑战。为了扩大我们的活体成像方法的范围，我们将采用和评估新型荧光“智能探针”。这些智能探针已在爱丁堡使用，通过尖端光纤显微镜对人类肺部的关键生物和病理事件进行成像，从而提供从计划的斑马鱼和小鼠研究到人类生命中患病器官的直接转化途径（该技术很容易应用于消化道和生殖系统）。我们的团队已经在使用各种模型动物方面建立了良好的记录，这些模型动物在关键靶细胞/器官中表达感兴趣的荧光标记分子。这些专业知识将保证我们开发出稳健的方法，重点关注具有科学挑战性的问题，例如：巨噬细胞在乳腺癌的发展中有何作用？神经元和神经胶质细胞在神经系统的发育、功能和修复中如何相互作用？性类固醇激素如何改变子宫内的细胞行为以及如何对其进行调节？与患有脑部疾病的模型小鼠相比，正常小鼠大脑中的中枢神经元蛋白质的分布如何？围绕活体成像在生物医学研究中的应用而令人兴奋的原因是它能够跟踪活组织内的细胞及其内容物，为研究蛋白质分布、分子相互作用和组织组成的动态变化提供了一个新的平台。目前，这种观察只能在组织内的单个位置进行——我们的目标是将数据采集扩展到整个器官，例如器官。询问小鼠大脑中突触蛋白的分布。为了实现这一目标，我们将把我们的活体技术与快速共焦显微镜上的系统 3D 图像采集相结合，能够扫描整个切片器官。由此产生的一系列 3D 图像将被存档，以确保数据保存，并将通过可搜索的基于网络的器官库提供给更广泛的研究界。这些详细描述特定标记蛋白质位置的分子“器官图”将使我们能够改进后续的“聚焦”功能性活体成像，并提供具有活体标本功能测量的高内涵 3D 器官图。这些创新研究将为我们的转化研究管道提供数据，并为改进健康问题的诊断和治疗方法的开发提供信息，这些问题包括癌症、多发性硬化症、不孕不育、心血管和 神经退行性疾病。

项目成果

Tumor initiating cells induce Cxcr4-mediated infiltration of pro-tumoral macrophages into the brain.

• DOI: 10.7554/elife.31918
• 发表时间: 2018-02-21
• 期刊: eLife
• 影响因子: 7.7
• 作者: Chia K;Mazzolini J;Mione M;Sieger D
• 通讯作者: Sieger D

In vivo subcellular resolution optical imaging in the lung reveals early metastatic proliferation and motility.

• DOI: 10.1080/21659087.2015.1086613
• 发表时间: 2015-09
• 期刊: Intravital
• 作者: Entenberg D;Rodriguez-Tirado C;Kato Y;Kitamura T;Pollard JW;Condeelis J
• 通讯作者: Condeelis J

Precise spatio-temporal control of rapid optogenetic cell ablation with mem-KillerRed in Zebrafish.

• DOI: 10.1038/s41598-017-05028-2
• 发表时间: 2017-07-11
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Buckley C;Carvalho MT;Young LK;Rider SA;McFadden C;Berlage C;Verdon RF;Taylor JM;Girkin JM;Mullins JJ
• 通讯作者: Mullins JJ

A Zebrafish Live Imaging Model Reveals Differential Responses of Microglia Toward Glioblastoma Cells In Vivo.

• DOI: 10.1089/zeb.2016.1339
• 发表时间: 2016-12
• 期刊: Zebrafish
• 影响因子: 2
• 作者: Hamilton L;Astell KR;Velikova G;Sieger D
• 通讯作者: Sieger D