Collaborative Research: Multimode Adaptive 3D Microscopy for Quantitative Analysis of Live-Cellular Dynamic Processes in Thick Samples

该项目的目标是设计、开发和研究一种计算光学显微镜系统，以生成分辨率大大提高的三维(3D)图像。该系统将使生物学家能够可视化和测量厚重样本中的活细胞动态过程，否则可能无法检测到这些过程，因为在传统显微镜中，随着成像深度的增加，像差会恶化。新系统将克服这些限制，同时在功能和分辨率方面提供更多好处。例如，可以同时提供高精度的荧光图像和3D位置信息，以及相位(折射率)和幅度(吸收)信息，从而每个生物特征将与3D空间中的定量信息相关联。该系统的另一项能力将使准确跟踪移动的微生物，甚至测量动态过程的速度。获得高深度精度的3D信息的前景为解决新的生物学应用开辟了无数可能性，例如研究上皮伤口愈合机制和癌细胞的形态动力学变化。该系统可用于检测培养细胞或肺组织中细胞迁移过程中细胞骨架蛋白(如肌动蛋白)的重塑。该系统专为各种成像条件下的优化性能而设计，将在许多生物和医疗应用中提供重大进展，这些应用严重依赖3D分辨率，而这是当前系统所不具备的。在生物仪器开发的多学科领域，将为不同的青年科学家和工程师提供培训机会。孟菲斯大学将建立一个新的教育、研究和跨学科工作实验室。从研究中获得的知识将成为孟菲斯大学计算成像新研究生课程的一部分，以及科罗拉多大学新的教学实验室模块的一部分。新的多模式显微镜将提供给两个校区的其他生物合作者。该项目还将利用与生物成像公司的协同合作伙伴关系，将新发现转化为工业应用。

The goal of this project is to design, develop and investigate a computational optical microscopy system to generate three-dimensional (3D) images with greatly enhanced resolution. The system will enable biologists to visualize and measure live-cellular dynamic processes in thick samples that might otherwise go undetected due to aberrations that worsen with imaging depth in conventional microscopes. The new system will overcome these limitations while providing additional benefits in terms of functionality and resolution. For example, it will be possible to simultaneously provide high accuracy fluorescent images and 3D position information, as well as phase (refractive index) and amplitude (absorption) information, so that each biological feature will be associated with quantitative information in 3D space. Another capability of the system will enable tracking moving micro-organisms accurately and even measuring the velocity of dynamic processes. The prospect of obtaining 3D information with high depth accuracy opens up myriad possibilities to address new biological applications such as studies of epithelial wound healing mechanisms and morphodynamic changes in cancer cells. The system could prove useful to detect the remodeling of cytoskeletal proteins, such as actin, during cell migration in culture or in lung tissue. The system, designed for optimized performance with respect to various imaging conditions, will provide significant advances in many biological and medical applications that critically depend on 3D resolution that is not available with current systems. Training opportunities will open for a diverse cadre of young scientists and engineers in the multi- disciplinary field of biological instrument development. A new laboratory for education, research, and interdisciplinary work at the University of Memphis will be established. Knowledge gained from the research will become part of a new graduate course in Computational Imaging at the University of Memphis and new teaching lab modules at the University of Colorado. The new multimodal microscope will be made available to other biological collaborators in both campuses.The project will also take advantage of synergistic partnerships with bio-imaging companies to transfer new discoveries into industrial applications.