Upgrades to an advanced multiphoton microscope with unique equipment to improve tissue access for intravital imaging and improved image stability

活体(活组织)成像是一种强大的工具，可以直接显示细胞及其在适当的解剖学背景下的行为。这项技术在细胞生物学、免疫学、神经科学和其他细胞生理学研究的不同领域有许多应用。这项应用是为了支持对现有多光子共聚焦显微镜进行两次升级，以克服成像活组织所固有的一些挑战。呼吸等生理过程引起的组织运动仍然是活体显微镜的技术障碍。为了解决这一问题，我们建议获得一种图像稳定三脚架，它可以物理地移动显微镜物镜，以匹配组织的生物运动。活体显微镜面临的另一个挑战是获取组织本身。与典型的小动物模型相比，标准多光子显微镜物镜较大(例如。(小鼠和大鼠)，有时需要做大手术才能腾出足够的空间。为了解决这个问题，我们建议购买针状客观内窥镜，它使用一个非常薄(&lt；1 mm)的旋转探头作为潜望镜。这将允许通过最小的手术接触到深层或其他无法触及的组织。总而言之，这些升级将大大增强现有显微镜的能力，提高使用它进行研究的效率和类型，并扩大用户基础和能够接触到这一重要成像技术的受训人员数量。

Intravital (live tissue) imaging is a powerful tool to directly visualize cells and their behaviour within their appropriate anatomical context. This technology has many applications to different fields of cell biology, immunology, neuroscience, and other investigations of cell physiology. This application is to support the acquisition of two upgrades to an existing multiphoton confocal microscope that will overcome some of the challenges inherent to imaging live tissues. Tissue movement due to physiological processes, such as breathing, remains a technical hurdle to intravital microscopy. To address this, we propose to acquire an Image Stabilization Tripod that physically moves the microscope objective to match the biological movement of the tissue. Another challenge to intravital microscopy is access to the tissue itself. Standard multiphoton microscope objectives are large compared to typical small animal models (ex. mice and rats), and sometimes extensive surgery is required in order to make sufficient space. To address this issue, we propose to acquire a Needle Objective Endoscope, that uses a very thin (<1mm), rotating probe as a periscope. This will allow access to deep or otherwise inaccessible tissues with minimal surgery. Together, these upgrades will considerably enhance the capabilities of the existing microscope, increase the efficiency and types of studies that can be performed using it, and broaden the user base and the number of trainees that have access to this important imaging technology.