An In Vivo/In Vitro 2-Photon Uncaging/Imaging Microscope

描述（由申请人提供）：双光子激发提供了优于其他类型的基于显微镜的成像的显著优势，特别是对于活体内或厚切片中的活细胞成像。事实上，在过去几年中，双光子成像的基础技术已经迅速发展。该提案的目的是为不断增长的麻省理工学院神经科学社区提供使用该技术的机会。该仪器将使研究人员能够在实验中同时对两种或更多种荧光蛋白进行成像，将扫描与复杂的技术相结合，包括电生理记录，刺激含有通道视紫红质或盐视紫红质的细胞，或快速释放生物活性分子。获得这些尖端技术将加强各种各样的研究计划，并促进NIH资助的研究的广泛阵列，包括解决以下问题的研究：发育中大脑突触形成的分子调节，控制成年大脑中新生神经元迁移和整合的因素，C损伤后的神经突再生。elegans，恐惧学习和消退后的结构变化，控制抑制性突触形成和稳定的因素，蛋白质错误折叠在帕金森病和朊病毒病小鼠模型进展中的作用，调节脑室发育的因素，抑制性中间神经元和星形胶质细胞在视觉皮层动力学中的作用的研究，自闭症和精神分裂症小鼠模型中的脊柱动力学，以及突触后支架蛋白对突触可塑性的贡献。该仪器还将用于进一步推进双光子成像技术，使旨在开发用于快速双光子激活细胞和快速读出神经活动的软件扫描协议的研究成为可能，特别是促进开发用于在成像过程中快速识别细胞的算法。

DESCRIPTION (provided by applicant): Two photon excitation offers significant advantages over other types of microscopy-based imaging, especially for imaging living cells either in vivo or in thick slices. Indeed, the technology underlying two photon imaging has rapidly advanced over the last few years. The objective of this proposal is to provide access to this technology to the growing MIT Neuroscience community. This instrument will enable researchers to perform simultaneous imaging of two or more fluorescent proteins in experiments combining scanning with sophisticated techniques, including electrophysiological recording, stimulation of cells containing channelrhodopsins or halorhodopsins, or rapid uncaging of bioactive molecules. Access to these cutting-edge techniques will enhance a broad variety of research programs, and facilitate an expansive array of NIH-funded research, including studies addressing the following: molecular regulation of synapse formation in the developing brain, factors controlling the migration and integration of newly born neurons in the adult brain, neurite regeneration following injury in C. elegans, structural changes following fear learning and extinction, factors governing inhibitory synapse formation and stabilization, the role of protein misfolding in the progression of mouse models of Parkinson's and prion disease, factors regulating the development of brain ventricles, studies of the role of inhibitory interneurons and astrocytes in visual cortex dynamics, spine dynamics in mouse models of autism and schizophrenia, and the contribution of postsynaptic scaffolding proteins to synaptic plasticity. This instrument will also be used to further the advancement of two photon imaging technologies, by enabling research aimed at developing software scanning protocols for rapid two-photon activation of cells and rapid readout of neural activity, and particularly facilitating the development of algorithms for rapid identification of cells during the imaging process.