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

抽象/项目总结

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.