Quantitative Multidimensional Imaging: A Centre of Excellence for Fluorescence Lifetime Imaging Microscopy

自80年代和90年代商业化以来，共聚焦和多光子显微镜已分别成为生物成像的支柱。随着它们变得更加成熟，生物研究人员可用的复杂程度已经发展到甚至被认为是显微镜最前沿的实验技术（如超分辨率显微镜和体内成像）也变得相对简单。尽管如此，荧光寿命成像显微镜等功能成像方法在专业实验室之外仍然具有挑战性。荧光寿命成像（FLIM）是用于测量生物细胞和组织中局部变化的关键显微技术。它被用作一种图像对比机制，用于报告细胞环境中的许多特征，对于理解生物系统的复杂性至关重要- King's目前在该领域处于世界领先地位。不幸的是，由于技术和分析程序的复杂性以及实施测定的开发时间，FLIM采用的障碍很高。为了保持在技术开发和生物成像之间的接口研究的前沿，我们认识到，这些设施必须公平地提供给该领域的所有研究人员。为了实现这一目标，我们建议购买一台Leica Stellaris 8 Dive Falcon，在King's显微镜创新中心内创建一个荧光寿命成像卓越中心。Falcon代表了FLIM、多光子和共焦成像的当前商业最新技术水平，非常适合支持拟议的科学，并将对MIC内发生的尖端发展具有高度互补性，包括高光谱FLIM、光片、超分辨率和高内容筛选（波兰，Ameer-Beg）。通过将Falcon与这些先进的成像平台共同放置在一个开放访问、集中管理的核心设施中，我们的目标是在更广泛的生物样本基础上普及FLIM的使用，并鼓励在多模式成像管道中使用这种强大的技术，以了解支持疾病的分子相互作用。（基础与生物医学科学，癌症与制药，口腔与颅面，心血管医学，免疫学与微生物）和国王学院的四个学院，提议利用这项技术的研究包括广泛的生物应用，包括癌症，神经科学，免疫学和解剖学。我们已经选择了几个项目，跨越广泛的研究，这将有助于通过建立这一系统。我们在King's和伦敦更广泛的用户群的实力和深度相当可观，将进一步加强King's的EuroBioImaging UK节点，以便从欧洲各地访问。

Confocal and Multiphoton microscopy have become the mainstay of biological imaging since their commercial introduction in the 80's and 90's respectively. As they have become more established the level of sophistication available to biological researchers has grown to the extent that even experimental techniques considered at the extreme forefront of microscopy (such as super-resolution microscopy and in vivo imaging) have become relatively straightforward. Despite this, functional imaging methods such as fluorescence lifetime imaging microscopy remain challenging outside specialist labs. Fluorescence lifetime imaging (FLIM) is a key microscopy technique used to measure local variations in biological cells and tissues. It is used as an image contrast mechanism to report on many characteristics within the cellular environment and are essential to understanding the complexities of biological systems - King's is currently world leading in this area. Unfortunately, the barriers for FLIM adoption are high due to the complexity of both the technology and analysis procedures and, development time to implement assays. To remain at the forefront of research at the interface between technology development and bioimaging we recognise that these facilities must be equitably available to all researchers in the field. To achieve this, we propose to purchase a Leica Stellaris 8 Dive Falcon to create a centre of excellence for fluorescence lifetime imaging to be located within the Microscope Innovation Centre at King's. The Falcon represents the current commercial state-of-the art for FLIM, multiphoton and confocal imaging, is highly appropriate to support the proposed science and will be highly complementary to cutting-edge developments occurring within the MIC including hyper-spectral FLIM, light-sheet, super-resolution and high content screening (Poland, Ameer-Beg). By co-locating the Falcon with these advanced imaging platforms in an open access, centrally managed core facility we aim to democratise the use of FLIM across a far broader base of biological samples and encourage the use of this powerful technique in multimodal imaging pipelines to understand molecular interactions underpinning disease.Extending across several schools (Basic & Biomedical Sciences, Cancer & Pharmaceutical, Oral & Craniofacial, Cardiovascular Medicine, Immunology & Microbial) and four faculties at King's, the research proposed to utilize this technology encompasses a wide range of biological applications including cancer, neuroscience, immunology and anatomy. We have selected several projects that span the broad spectrum of research which will be facilitated through the establishment of this system. The strength and depth in our broader user base across King's and across London is considerable and will further strengthen the EuroBioImaging UK Node at King's to enable access from across Europe.