A multiphoton system that allows simultaneous multiphoton imaging and 3D optical stimulation

摘要/项目摘要 在该项目中，我们申请资金购买直立式布鲁克NeuraLight 3D空间激光调制器（SLM）Ultima 多光子显微镜，使同时多光子成像和3D全息光学刺激。 该仪器将允许神经元活动的同时深部脑，解耦，双光子功能成像 和SLM全息光遗传学刺激来研究和理解神经连接以及神经如何 网络控制行为。该仪器将被安置在新泽西儿童健康研究所（JIU）， 罗格斯大学罗伯特伍德约翰逊医学院（RWJMS）支持研究， 突触和细胞机制的精神障碍的细胞类型和电路的具体方式。这 仪器将是罗格斯大学生物医学和健康科学（RBHS）和罗格斯大学新的 Brunswick/皮斯卡特维校区，并将填补多个研究人员可用技术的关键空白 由罗格斯大学的NIMH资助。该系统的核心用户将优先考虑NIMH资助的研究实验室 他擅长开发新型光学传感器，研究恐惧、抑郁、自闭症和妥瑞症 综合征，但也将提供给其他NIH资助的研究人员使用。这种尖端仪器 将满足对细胞类型和电路的深层组织成像的急剧增长的需求的关键需求- 特定的光遗传学操作，对于理解复杂的大脑疾病和行为至关重要， 目前校园内的任何其他系统都无法满足。光学技术的最新发展， 特异性视蛋白和遗传编码的神经元活性报告物，包括GCaMPs（钙）、Voltron & ASAP （电压）、dLight或GRAB-DA（多巴胺），以及用于由GPCR介导的传递的报告基因（RTGR） 我们目前正在开发的，使这些研究范式成为可能。重要的是我们有 获得了来自CNOJ，Rutgers-RWJMS和Rutgers-School of Arts and Sciences的坚定承诺， 本申请获得这种最先进的直立多光子显微镜系统，包括指定的 空间，额外仪器成本的资金，以及对延长服务合同的长期支持 系统的维护。主要研究者和一名高级实验室研究助理，沿着当地 技术专家，将负责所有用户培训和日常维护，并为用户提供 协助、实验设计咨询以及系统配置和最佳系统使用建议。 成立了一个由具有生物光子学专业知识的成员组成的咨询委员会，以确保 正确和有效地使用设施。我们预计，这一系统的加入不仅将大大 加强目前NIMH资助的项目，但也将允许用户探索新的问题，以前没有 由于缺乏能够以高速神经元活动深入成像到样品中的平台， 操纵因此，该仪器将在促进神经科学新发现方面发挥关键作用 并帮助设计治疗精神疾病的新疗法。

Abstract/Project Summary In this project, we request funds to acquire an upright Bruker NeuraLight 3D spatial laser modulator (SLM) Ultima multiphoton microscope to enable simultaneous multiphoton imaging and 3D holographic optical stimulation. The instrument will allow simultaneous deep brain, decoupled, two-photon functional imaging of neuronal activity and SLM holographic optogenetic stimulations to study and understand neural connectivity and how neural networks control behavior. The instrument will be housed at the Child Health Institute of New Jersey (CHINJ) at Rutgers Robert Wood Johnson Medical School (RWJMS) to support research in unraveling the molecular, synaptic, and cellular mechanisms underlying mental disorders in a cell type- and circuity-specific manner. This instrument will be the first of its kind on the Rutgers Biomedical and Health Sciences (RBHS) and Rutgers New Brunswick/Piscataway campuses and will fill a critical void in available technologies to multiple investigators funded by NIMH at Rutgers. The core users of the system will be prioritized to NIMH-funded research laboratories with expertise in developing novel optical sensors, and studying fear, depression, and autism and Tourette syndromes, but will also be available for use by other NIH-funded researchers. This cutting-edge instrumentation will meet a critical need in the dramatically increased demand for deep tissue imaging with cell type and circuitry- specific optogenetic manipulations, essential for understanding complex brain disorders and behaviors, which cannot be met by any other systems currently available on campus. Recent developments in optics technologies, specific opsins and genetically encoded neuronal activity reporters including GCaMPs (calcium), Voltron & ASAP (voltage), dLight or GRAB-DA (dopamine), as well as Reporters for Transmission mediated by GPCRs (RTGRs) which we are currently developing, have made these research paradigms possible. Importantly, we have obtained strong commitments from the CHINJ, Rutgers-RWJMS, and Rutgers-School of Arts and Sciences for this application to acquire this state-of-the-art upright multiphoton microscopy system, including designated space, funding for additional instrumentation costs, and long-term support for an extended service contract and maintenance for the system. The principal investigator and a senior lab research associate, along with local technical experts, will be responsible for all user training and routine daily maintenance, as well as providing user assistance, consultation on experimental design and advice on system configuration and optimal system use. An advisory committee composed of members with strong expertise in biophotonics has been formed to ensure proper and efficient use of the facility. We anticipate that the addition of this system will not only greatly strengthen current NIMH-funded projects but will also allow users to explore new questions previously not addressable due to the absence of a platform able to image deeply into samples with high speed neuronal activity manipulations. Therefore, this instrument will play a critical role in promoting new discoveries in neuroscience and help towards devising novel therapeutics for treating mental disorders.