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

描述(申请人提供)：该项目旨在开发猎户座显微镜系统，这是一种高度创新的软硬件结合产品，将促进使用延时视频显微镜(TLVM)自动研究神经元前体迁移动力学和体外轴突生长。为了实现这一目标，猎户座显微镜系统将把新的软件与MBF最近开发并获得专利的两个创新的显微镜硬件设备结合在一起：(I)可移动的扩展显微镜物镜设备，以及(Ii)双目标显微镜设置。猎户座显微镜系统将首次允许研究人员用两个显微镜物镜同时检查组织培养物(使用荧光照明)，并有可能在XYZ方向上相互独立地移动物镜。这一系统将有助于发现有关神经元前体和轴突生长的新发现。哺乳动物大脑的发育依赖于神经元前体的广泛迁移。有必要更好地了解发育中大脑中神经元前体迁移的调节(和纠正)，以便开发新的方法来预防和治疗各种神经和神经精神疾病，如儿童期发作性癫痫、自闭症、精神分裂症、注意缺陷多动障碍、无脑以及HIV-1感染儿童的神经学和神经行为后遗症。此外，更好地了解轴突生长的动力学(特别是纠正受损的轴突生长)已成为开发新的治疗策略以对抗阿尔茨海默病、帕金森病、亨廷顿病和神经艾滋病等神经退行性疾病的重要方面。为了研究神经元前体迁移和轴突生长的动力学，研究人员通常在体外分离的神经元培养物或器官型脑片培养物上进行TLVM，检查单一视野。存在几种软件和软件/硬件组合产品来协助调查人员进行这类研究。然而，这些产品都不包含以下功能，这些功能在高级自动化分析神经元前体迁移和轴突生长的动力学中被认为是至关重要的：(I)识别和跟踪迁移的神经元前体和生长的轴突，在低倍放大的大组织培养中的许多微观视野中，同时，(Ii)确定已确定的迁移的神经元前体的不同迁移模式或在高倍镜下生长的神经突起的细微变化。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，对神经科学研究和开发预防或抗击上述疾病的新治疗策略具有很高的相关性。因此，猎户座显微镜系统的发展代表着超越最先进水平的明显进步，为神经科学研究界和整个社会带来了巨大的好处。 公共卫生相关性：拟议的猎户座显微镜系统将在基础神经科学研究以及药理学和生物技术研究和开发方面开辟新的视野，通过提供一个强大的自动化双目标显微镜系统来研究神经元前体的迁移动力学和培养皿中神经元突起的生长。该项目的总体效果将有助于研究新的治疗策略，以预防和治疗各种神经、神经精神和神经退行性疾病，如儿童期发作的癫痫、自闭症、精神分裂症、注意力缺陷多动障碍、无脑畸形、以及感染艾滋病毒的儿童的神经和神经行为后遗症，以及阿尔茨海默病、帕金森氏病、亨廷顿病和神经艾滋病。

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. PUBLIC HEALTH RELEVANCE: The proposed Orion microscope system will open new horizons in basic neuroscience research as well as in pharmacological and biotechnological research and development, by providing a powerful, automated dual objective microscope system to investigate dynamics of migration of neuronal precursors and outgrowth of neuronal processes in a petri dish. The overall effect of this project will benefit research in novel therapeutic strategies for preventing and treating various neurological, neuropsychiatric, and neurodegenerative 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, as well as Alzheimer's disease, Parkinson's disease, Huntington's disease, and NeuroAIDS.