Multi-Photon Laser Scanning Microscope

炎症过程与多种疾病有关，如感染、癌症转移、血栓形成和缺血性疾病。然而，对潜在病理生理的理解仍然不完整。详细阐明体内先天和适应性免疫反应，特别是免疫细胞募集、迁移、激活及其在不同生态位和器官环境中的功能，对于开发解决组织炎症的新治疗策略至关重要。多光子活体显微镜彻底改变了高分辨率成像，使研究人员能够深入了解体内组织和器官系统内的炎症过程。这是通过实时和长期高分辨率深部组织成像实现的，同时仅施加最小的光毒性。使用多光子活体显微镜结合转基因小鼠模型是我们研究癌症转移，血栓形成和免疫细胞运输及其在无菌炎症和感染中的功能的基石。本提案中概述的所有研究项目都依赖于多光子活体显微术来实现其目标。当我们与活体动物一起工作时，低光毒性的快速深层组织成像是一个关键问题。此次收购的先进多光子显微镜将提供最先进的功能，包括高时空分辨率和高灵活性的扫描策略，如灵敏度和速度。我们目前的多光子活体显微镜设备已经使用了12年，并以最大容量运行。然而，该系统缺乏灵敏度、速度、分辨率、激光激发能力以及机动平台。总之，目前的系统在速度和分辨率方面没有提供最先进的成像能力，而且还禁止在免疫生物学研究中扩展边界，例如通过体内照片处理。获得一个新的集成多光子系统将使我们的研究项目保持竞争力，并开展尖端的科学研究。

Inflammatory processes are associated with a broad variety of conditions such as infections, cancer metastasis, thrombosis and ischemic diseases. However, understanding of the underlying pathophysiology has remained incomplete. Detailed elucidation of innate and adaptive immune responses in vivo, specifically immune cell recruitment, migration, activation and their functions within distinct niches and organ environments, is of pivotal importance for the development of novel therapeutic strategies to resolve tissue inflammation. Multiphoton intravital microscopy has revolutionized high-resolution imaging allowing researchers to gain profound insight into inflammatory processes within tissues and organ systems in vivo. This is achieved by enabling real-time and long-term high-resolution deep tissue imaging, while exerting only minimal phototoxicity. The use of multiphoton intravital microscopy in combination with transgenic mouse models represents a cornerstone of our research into cancer metastasis, thrombosis and immune cell trafficking and functions in sterile inflammation and infections.All research projects outlined in this proposal rely on multiphoton intravital microscopy to achieve their aims. As we work with live animals, fast deep tissue imaging with low photo-toxicity is a critical issue. The proposed acquisition of an advanced multiphoton microscope would provide state-of-the-art functionality including high spatio-temporal resolution combined with high flexibility in scanning strategies, such as sensitivity and speed. Our current multiphoton intravital microscope device is 12 years old and running at maximum capacity. However, the system lacks sensitivity, speed, resolution, laser excitation capabilities, as well as a motorized stage. Together, the current system does not provide state-of-the-art imaging capabilities in terms of speed and resolution but also prohibits expansion of boundaries in the study of immunobiology, for example through in vivo photo-manipulation. The acquisition of a new integrated multiphoton system will allow our research projects to stay competitive and carry out cutting edge scientific research.