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

描述(申请人提供)：与其他类型的基于显微镜的成像相比，双光子激发具有显著的优势，特别是对于活体或厚切片中的活细胞进行成像。事实上，双光子成像背后的技术在过去几年里迅速发展。这项提议的目的是为不断增长的麻省理工学院神经科学界提供使用这项技术的途径。该仪器将使研究人员能够在结合扫描和尖端技术的实验中同时对两种或两种以上的荧光蛋白质进行成像，包括电生理记录、刺激含有通道的视紫红质或卤视紫质的细胞，或快速去除生物活性分子。获得这些尖端技术将加强各种各样的研究项目，并促进由美国国立卫生研究院资助的大量研究，包括以下研究：发育中大脑中突触形成的分子调控，控制成年大脑中新生神经元迁移和整合的因素，线虫损伤后的轴突再生，恐惧学习和消失后的结构变化，抑制突触形成和稳定的因素，蛋白质错误折叠在帕金森和普里恩疾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质动力学中的作用研究，自闭症和精神分裂症小鼠模型的脊柱动力学，以及突触后支架蛋白对突触可塑性的贡献。该仪器还将用于促进双光子成像技术的进步，使旨在开发软件扫描协议的研究成为可能，以便快速激活细胞和快速读出神经活动，特别是促进开发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.