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

共焦显微镜和多光子显微镜自80年代S和90年代S商业化引进以来，已成为生物成像的主流。随着它们变得更加成熟，生物研究人员可以获得的复杂程度已经增长到即使是被认为是显微镜最前沿的实验技术(如超分辨率显微镜和活体成像)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。荧光寿命成像是一项关键的显微技术，用于测量生物细胞和组织的局部变异。它被用作一种图像对比度机制，用于报告细胞环境中的许多特征，对于了解生物系统的复杂性至关重要-King‘s目前在这一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检测的开发时间，采用FILM的门槛很高。为了在技术开发和生物成像之间的界面上保持在研究的前沿，我们认识到这些设施必须公平地向该领域的所有研究人员提供。为了实现这一目标，我们建议购买徕卡Stellaris 8 Dive Falcon，以创建一个卓越的荧光寿命成像中心，位于国王的显微镜创新中心。Falcon代表了当前薄膜、多光子和共焦成像的商业最先进水平，非常适合支持拟议的科学，并将与MIC内发生的尖端发展高度互补，包括高光谱薄膜、光片、超分辨率和高含量筛选(波兰，Ameer-Beg)。通过将猎鹰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一强大的技术，以了解支撑疾病的分子相互作用。这项研究扩展到几个学院(基础和生物医学、癌症和制药、口腔和颅面、心血管内科、免疫学和微生物)和国王学院的四个学院，建议利用这项技术的研究涵盖广泛的生物应用，包括癌症、神经科学、免疫学和解剖学。我们选择了几个研究范围广泛的项目，这些项目将通过这一系统的建立而得到便利。我们在King‘s和整个伦敦的更广泛的用户基础的实力和深度是相当大的，并将进一步加强King’s的EuroBioImaging UK节点，使其能够从整个欧洲访问。

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.