"Establishing fast superresolution live cell and tissue imaging with photomanipulation"

人类免疫系统对活跃的健康和解决疾病至关重要，但信息缺失。近年来，免疫反应依赖于器官、免疫细胞和分子的多尺度关系，这一点在生物医学科学中得到了广泛的接受，但整体观点仍然难以捉摸。多年来，显微镜一直是免疫反应科学发现的来源。然而，直到最近，还没有一种具有适当灵敏度的显微镜技术来研究不同尺度的免疫反应。这一MRC中端设备应用的重点是这一挑战的核心，旨在建立具有定制光模拟的ZEISS Elyra 7 SIM 2，该光模拟可提供跨细胞和分子的活组织的快速定量超分辨率生物成像。ZEISS Elyra提供的具有光操纵的特殊光操纵功能允许光激活/光子跳跃/光漂白，从而跟踪周围组织环境中的特定分子和/或细胞。我们在多个场合测试了ZEISS Elyra 7，发现该仪器是解决所提出的研究问题的理想仪器，设计精良，用户友好，与使用定制SIM显微镜进行的实验直接比较。ZEISS Elyra 7在这方面尤其令人印象深刻，它允许人们轻松安装和定位样本，并在非常稳定的条件下保持SIM，适合扩展的单细胞和薄10-100微米组织切片生物成像。据我们所知，这将是英国唯一一个具有光操纵功能的ZEISS Elyra 7 SIM2平台。具有光操纵功能的Zeiss Elyra 7 SIM2满足了组织业界多年来的所有要求。具有光操控的ZEISS Elyra 7的用户基础来自MRC和Wellcome资助的研究人员联盟，并将在UKRI-MRC的职权范围内进行研究，重点是感染和免疫、全球健康、神经科学和精神健康、MRC对COVID的反应以及转化医学。ZEISS Elyra 7的MRC中端设备的成功应用将显著扩展牛津和英国生物成像的广度、容量和能力，使英国的生物成像能力保持在国际研究的前沿。

Human immune system is critical for active health and for resolving disease, but information is missing. Over recent years it has become widely accepted in the biomedical sciences that the immune response depends on the multiscale relationships of its organs, immune cells, and molecules, but a holistic perspective remains elusive. Microscopy has, for many years, been a source of scientific discovery for the immune response. However, until recently there has been no microscopy technology with the right sensitivity to study the immune response across scales. The focus of this MRC Mid-range Equipment application sits at the heart of this challenge aiming to establish the ZEISS Elyra 7 SIM 2 with custom-built photomanipuation, which offers fast quantitative superresolution bioimaging of living tissues across scales down to cells and molecules. The special photomanipulation feature provided by this ZEISS Elyra with photomanipulation allows to photoactivate/photouncage/photobleach and thus track specific molecules and/or cells in the surrounding tissue context.We tested the ZEISS Elyra 7 at multiple occasions and found the instrument to be ideal for the proposed research questions, well engineered, and user friendly in direct comparison with experiments acquired at bespoke SIM-based microscopes. The ZEISS Elyra 7 was particularly impressive in this respect, allowing one to easily mount and locate samples, as well as maintain SIM under very stable conditions suitable for extended single cell and thin 10-100um tissue slice bioimaging. To the best of our knowledge, this will be the only ZEISS Elyra 7 SIM 2 platform with photomanipulation in the UK.The Zeiss Elyra 7 SIM2 with photomanipulation meets all the requirements demanded by the tissue community for years. The user base for the ZEISS Elyra 7 with photomanipulation stems from a consortium of MRC-funded and Wellcome-funded investigators and will be undertaking research within UKRI-MRC remit, focusing on infections and immunity, global health, neuroscience and mental health, MRC response to COVID, and translational medicine.A successful MRC Mid-range Equipment application for the ZEISS Elyra 7 would significantly expand the breadth, capacity, and capabilities of bioimaging in Oxford and in the UK, keeping UK's bioimaging abilities at the forefront of international research.