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基本技术，并且有助于解开控制组织发育、分化和驱动疾病过程的生物系统的复杂性。我们的方法是将联合收割机无标记相干拉曼成像模式与双光子荧光成像模式相结合，该模式使用给定化学品的拉曼活性振动频率产生图像对比度，以提供对亚细胞和组织组织的洞察。双正交化学的最新进展允许小药物分子的拉曼标记和随后的可视化，自2011年我们建立了目前的多模态成像平台以来，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成像能力来显著增强和扩展成像能力。我们的测量的稳定性、灵敏度、准确度和速度，同时还能够实现远红荧光团的成像，由于更高波长到组织中的增强穿透，远红荧光团在体内探针的设计中变得越来越流行。新的设置将提供一个独特的平台，在活体动物（包括小鼠和斑马鱼）中进行活体成像，为基本生物过程的空间和时间控制提供无与伦比的见解。此外，获得汇集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影响。这将支持科学进步，并使技术创新，将影响生物科学的所有领域。该提案的目的是提供一个国家的最先进的多模态成像平台提供给整个UoE的科学界，其他学术机构在英国，和我们的合作伙伴在行业。该提案汇集了具有生物成像专业知识的多学科研究人员（化学家，生物学家，工程师），具有广泛的多样化研究目标。因此，新平台将支持BBSRC未来十年几个关键战略优先事项的科学研究：对抗微生物耐药性;使用动物进行研究的3R;可持续地提高农业生产;生物科学的系统方法;生物科学的技术开发;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.