System for multidimensional analysis of growth and migration of neurons and neuro

描述（申请人提供）：本项目旨在开发Orion显微镜系统，这是一种高度创新的软件/硬件组合产品，将有助于使用延时视频显微镜（TLVM）在体外自动研究神经元前体和神经突生长的动态迁移。为了实现这一目标，Orion显微镜系统将新型软件与MBF最近开发并获得专利的两种创新显微镜硬件设备相结合：(i)可移动扩展显微镜物镜设备，（ii）双物镜装置。猎户座显微镜系统将允许研究人员首次同时使用两个显微镜物镜（使用荧光照明）检查组织培养物，并且可以在XYZ方向上相互独立地移动物镜。该系统将有助于发现有关神经元前体和神经突生长的新发现。哺乳动物大脑的发育依赖于神经元前体的广泛迁移。为了开发预防和治疗各种神经和神经精神疾病的新方法，如儿童期癫痫、自闭症、精神分裂症、注意缺陷多动障碍、缺脑畸形，以及HIV-1感染儿童的神经和神经行为后遗症，需要更好地了解发育中的大脑中神经元前体迁移的调节（和纠正干扰）。此外，更好地了解神经突生长的动力学（特别是受损神经突生长的矫正）已经成为开发新的治疗策略以对抗神经退行性疾病（如阿尔茨海默病、帕金森病、亨廷顿病和神经艾滋病）的重要方面。为了研究神经元前体迁移和神经突生长的动力学，研究人员通常对离体神经元培养物或体外脑器官型切片培养物进行TLVM，检查单个视野。存在一些软件和组合软件/硬件产品来协助研究者进行此类研究。然而，这些产品都不包括以下功能，这些功能被认为是神经前体和神经突生长迁移动态的高级自动化分析的关键：(i)在低放大率下，在大型组织培养的许多显微镜视野中识别和跟踪迁移的神经元前体和生长的神经突，同时，（ii）确定已识别的迁移神经元前体的不同迁移模式或在高倍率下生长的神经突的细微变化。这将为理解神经元在大脑发育、重组和退化过程中的动态行为开辟新的视野，对神经科学研究和预防或对抗上述疾病的新治疗策略的发展具有高度相关性。因此，猎户座显微镜系统的发展代表了超越最先进技术的明显进步，对神经科学研究界和整个社会都有很大的好处。

DESCRIPTION (provided by applicant): This project aims at developing the Orion microscope system, a highly innovative combined software/hardware product, which will facilitate automated investigations of dynamics of migration of neuronal precursors and neurite outgrowth in vitro using time-lapse video-microscopy (TLVM). To achieve this goal, the Orion microscope system will combine novel software with two innovative microscope hardware devices that were recently developed and patented by MBF: (i) movable Extended Microscope Objective devices, and (ii) a dual- objective microscope setup. The Orion microscope system will allow investigators, for the first time, to inspect a tissue culture with two microscope objectives simultaneously (using fluorescence illumination), with the possibility of moving the objectives in XYZ directions independent of each other. This system will help uncover new discoveries about neuronal precursors and neurite outgrowth. The development of the mammalian brain is dependent on extensive migration of neuronal precursors. There is need for a better understanding of the regulation of (and correction of disturbed) migration of neuronal precursors in the developing brain, in order to develop novel approaches for preventing and treating various neurological and neuropsychiatric disorders such as childhood-onset epilepsy, autism, schizophrenia, attention deficit hyperactivity disorder, lissencephaly, and the neurologic and neurobehavioral sequelae seen in children with HIV-1 infection. Furthermore, better insights into the dynamics of neurite outgrowth (and particularly the correction of impaired neurite outgrowth) have become an important aspect in the development of novel therapeutic strategies to combat neurodegenerative disorders such as Alzheimer's disease, Parkinson's disease, Huntington's disease, and NeuroAIDS. To study the dynamics of neuronal precursor migration and neurite outgrowth, investigators usually perform TLVM on dissociated neuron cultures or organotypic slice cultures of the brain in vitro, inspecting single fields-of-view. Several software and combined software/hardware products exist to assist investigators in such studies. However, none of these products comprise the following functionality that is considered critical in advanced, automated analyses of the dynamics of migration of neuronal precursors and neurite outgrowth: (i) identifying and follow up migrating neuronal precursors and growing neurites across many microscopic fields-of-view within large tissue cultures at low magnification and, simultaneously, (ii) determining the different modes of migration of the identified migrating neuronal precursors or subtle changes in growing neurites at high magnification. This will open new horizons in understanding the dynamic behaviors of neurons during development, re-organization, and degeneration of the brain, with high relevance for both neuroscience research and the development of novel therapeutic strategies to prevent or combat the aforementioned disorders. Accordingly, the development of the Orion microscope system represents clear progress beyond the state-of-the-art, with great benefits for the neuroscience research community and society in general.