Acquisition of a Multiphoton Microscope for Cellular Programming

描述(由申请人提供)：这一定制多光子显微镜的共享仪器提案是宾夕法尼亚大学在细胞编程方面更大的机构努力的一部分。从战略上讲，这一努力位于宾夕法尼亚大学的五所学校-医学、牙科、兽医、艺术和科学以及工程和应用科学-的交叉点上。我们要求配备一台多光子显微镜，以便在细胞编程中执行三个独立但相关的工作。这些努力包括：在单细胞水平，其中我们使用由PI项目之一(Jim Eberwin，药理学，医学院)开发的新技术来可控地向活细胞传递限定的mRNA群体以改变其细胞表型，在多细胞水平，其中我们使用新型光聚合物配方来组装复杂的三维细胞培养底物(Chris Chen，生物工程，工程学院和应用科学)，具有可调节的微环境以建立血管组织和软骨，以及在组织规模，我们使用广泛可用的光学激活技术来研究皮质和海马区神经回路的体内编程，以了解在疾病或损伤期间发生的变化(David Meaney，生物工程、工程和应用科学学院)。这个新的显微镜系统将取代宾夕法尼亚大学工程综合体中现有的已有12年历史的BioRad多光子显微镜。目前的BioRad系统确实满足了上述应用的高技术要求。此外，宾夕法尼亚大学校园内没有广泛可用的现有系统来执行这项工作。因此，对这种显微镜系统有很大的需求。在一个显微镜平台上结合这三项“基础技术”可以显著推进广泛领域的研究课题，如细胞分化、再生医学以及神经疾病和神经行为的病因学。将两到三项基础技术整合到一个主题领域的潜力为生命系统如何形成和再生组织提供了几乎无限的前沿进展，并为组装新型组织替代或基于RNA的治疗方法开发了一个平台。

DESCRIPTION (provided by applicant): This shared instrumentation proposal for a customized multiphoton microscope is part of a larger institutional effort at Penn in cellular programming. Strategically, this effort lies at the intersection between five of Penn's schools - Medicine, Dental, Veterinary, Arts and Sciences, and Engineering and Applied Science. We request a multiphoton microscope equipped to perform three separate, but related efforts, in cellular programming. The efforts include: At the single cell level, where we use a novel technology developed by one of the project PIs (Jim Eberwine, Pharmacology, School of Medicine) to controllably deliver a define mRNA population to living cells to redirect their cellular phenotype, At the multicellular scale, where we use novel photopolymer formulations to assemble complex, three-dimensional cell culture substrates (Chris Chen, Bioengineering, School of Engineering and Applied Science) with tunable microenvironments for building vascularized tissue and cartilage, and At the tissue scale, where we use widely available optical activation techniques to study the in vivo programming of neural circuits in the cortex and hippocampus to understand changes that occur during disease or injury (David Meaney, Bioengineering, School of Engineering and Applied Science). This new microscope system will replace an existing 12 year old BioRad multiphoton microscope in the engineering complex at the University of Pennsylvania. The current BioRad system does meet the high technical demands of the above applications. Moreover, there is no widely available existing system on the Penn campus to perform this work. Therefore, there is substantial need for this microscope system. The combination of these three 'base technologies' on one microscope platform can significantly advance research topics in broadly diverse areas such as cellular differentiation, regenerative medicine, and the etiology of neurological disease and neurobehavior. The potential of integrating two or three of the base technologies into a single topic area provides nearly limitless possibilities for cutting edge advances in how living systems form and regenerate tissue, as well as developing a platform for assembling novel tissue replacement or RNA-based therapeutic approaches.