Multimodal coherent Raman and two-photon fluorescence imaging as a discovery tool in bioscience

生物成像是支撑生物科学的一项基础技术，在揭示控制组织发育、分化和驱动疾病过程的生物系统的复杂性方面发挥了重要作用。我们的方法是将无标记相干拉曼成像模式（利用给定化学物质的拉曼主动振动频率产生图像对比度）与双光子荧光成像模式相结合，以深入了解亚细胞和组织组织。双正交化学的最新进展使得小药物分子、药物输送载体和细胞成分的拉曼标记和随后的可视化得以实现，这使这项技术的好处扩展到更广泛的科学基础。自2011年我们建立目前的多模态成像平台以来，在仪器设计方面取得了一些进展，这意味着我们可以通过提高测量的稳定性、灵敏度、准确性和速度来显着增强和扩展成像能力，同时还可以对远红色荧光团进行成像，由于更高波长对组织的渗透增强，远红色荧光团在体内探针的设计中越来越受欢迎。新的装置将提供一个独特的平台，对包括老鼠和斑马鱼在内的活体动物进行活体成像，为基本生物过程的空间和时间控制提供无与伦比的洞察力。此外，获得提出该提案的研究人员联盟的专业知识和知识（他们拥有出色的出版物记录并获得包括BBSRC在内的财政支持）将推动成功并确保所进行的研究产生最大的影响。这将支持科学进步并使影响生物科学所有领域的技术创新成为可能。该提案的目的是为整个英国的科学界、整个英国的其他学术机构以及我们的行业合作伙伴提供一个最先进的多模态成像平台。该提案汇集了一个多学科的研究人员联盟（化学家、生物学家、工程师），他们在生物成像方面具有广泛的不同研究目标。因此，新平台将支持BBSRC未来十年的几个关键战略重点的科学研究：抗击微生物耐药性；3Rs在动物研究中；可持续提高农业生产水平；生物科学的系统方法；生物科学技术发展；合成生物学。

Bioimaging is a fundamental technology underpinning bioscience and has been instrumental in unravelling the complexities of biological systems that control tissue development, differentiation and that drive disease processes. Our approach has been to combine label-free coherent Raman imaging modalities, which generate image contrast using the Raman active vibrational frequency of a given chemical, with two-photon fluorescence imaging modalities to provide insight into both subcellular and tissue organisation. Recent advances in biorthogonal chemistry which allows Raman tagging and subsequent visualisation of small drug molecules, drug delivery vehicles and cellular constituents has extended the benefit of this technology to a much wider scientific base.Several advances in instrument design have been made since we set up our current multimodal imaging platform in 2011 which means that we can significantly enhance and extend the imaging capabilities by increasing the stability, sensitivity, accuracy and speed of our measurements, while also enabling imaging of far-red fluorophores which are becoming increasingly popular in the design of in vivo probes due to the enhanced penetration of higher wavelengths into tissue. The new set-up will provide a unique platform to carry out intravital imaging in live animals, including mice and zebrafish, providing unrivalled insight into spatial and temporal control of fundamental biological processes. In addition, access to the expertise and knowledge of the consortium of researchers that have put together this proposal (who have an excellent track record of publications and securing financial support including from the BBSRC) will drive success and ensure maximal impact of the research undertaken. This will support scientific advances and enable technological innovation that will impact all areas of the biological sciences.The aim of this proposal is to provide a state-of-the-art multimodal imaging platform available to scientific communities across the UoE, other academic institutions throughout the UK, and our partners in industry. The proposal brings together a multidisciplinary consortium of researchers (chemists, biologists, engineers) with expertise in bioimaging with a broad range of diverse research goals. The new platform will therefore underpin scientific research in several key strategic BBSRC priorities for the next decade: combatting microbial resistance; 3Rs in research using animals; sustainably enhancing agricultural production; systems approaches to the biosciences; technology development for the biosciences; synthetic biology.