Zeiss Axioimager Z2 Imaging System for Array Tomography

描述（由申请人提供）：突触是介导神经系统中神经元之间通信的细胞-细胞接触的专门位点。现在人们普遍认为，兴奋性或抑制性突触的异常发育或功能有助于神经功能障碍，如精神发育迟滞、自闭症谱系障碍和癫痫。为了了解突触功能障碍是如何成为这些神经系统疾病的基础的，我们最重要的是要发现突触在非扰动状态下是如何形成和发挥功能的。用户组中的五名研究人员的NIH赞助的研究特别关注这一研究目标：了解突触连接如何在各种实验生物的神经系统中形成，修改和维持。项目包括：中枢和外周神经系统中突触形成的机制、神经调节和内在兴奋性的调节。 阵列断层扫描是一种新的成像方式，代表了一种新的方法，在完整的神经系统突触结构的高分辨率成像。我们计划购买的Zeiss Axio Imager Z2荧光成像系统将允许该用户组使用斯坦福大学的Stephen Smith博士及其同事设计的创新切片和重建策略，以前所未有的分辨率研究突触结构和功能。由于在相对厚的组织切片（例如255 μ m）中的抗体穿透问题和在成像期间沿Z轴的有限分辨率（沿着Z轴），使用针对脑组织切片上的突触标记物的抗体的传统显微镜化学术产生突触点的差分辨率，即使当采用共聚焦显微镜时。阵列断层扫描通过对神经系统组织的冷冻切片（例如70 nm）进行抗体染色来规避这些问题。有许多额外的好处，利用阵列断层扫描超过传统的免疫组化，包括免疫荧光成像后，使用扫描电子显微镜从组织获得超微结构信息的可能性，也许最重要的是我们的目的，能够进行多轮的抗体染色的同一组织切片。当组织的位置固定在载玻片上时，用针对突触蛋白的不同抗体重复染色将允许编目单个神经元中的所有突触中存在或不存在哪些突触成分。用户组的所有五名研究人员都达到了用传统免疫荧光方法测定突触形态和功能的极限。因此，这是至关重要的，以促进这些群体的研究使命，阵列层析成像技术可在布兰迪斯校园。 公共卫生相关性：现在，许多研究指出，突触形成缺陷可能是自闭症、智力迟钝和癫痫等神经系统疾病的原因。了解异常突触形成如何导致这些广泛的神经损伤的一种方法是首先使用光学显微镜研究突触如何形成，维持和在非病理状态下发挥作用。收购Zeiss Axioimager Z2阵列断层扫描系统将允许前所未有的高分辨率成像突触从神经系统的各种实验生物。

DESCRIPTION (provided by applicant): Synapses are specialized sites of cell-cell contact that mediate communication between neurons in the nervous system. It is now widely believed that aberrant development or function of either excitatory or inhibitory synapses contributes to neurological impairments such as mental retardation, autism spectrum disorders and epilepsy. To understand how synapse dysfunction underlies these neurological disorders, it is paramount that we discover how synapses form and function in the non-perturbed state. The NIH-sponsored research of the five investigators in the user group is specifically focused on this research goal: understanding how synaptic connections are formed, modified, and maintained in the nervous system of a variety of experimental organisms. Projects include: mechanisms of synapse formation in the central and peripheral nervous system, neuromodulation, and the regulation of intrinsic excitability. Array tomography is a novel imaging modality that represents a new approach to high resolution imaging of synaptic structure in intact nervous systems. The Zeiss Axio Imager Z2 fluorescence imaging system that we propose to purchase will allow this user group to study synaptic structure and function at an unprecedented resolution using an innovative sectioning and reconstruction strategy devised by Dr. Stephen Smith and colleagues at Stanford University. Traditional immunohistochemsitry using antibodies against synaptic markers on brain tissue sections yields poor resolution of synaptic puncta due to antibody penetration problems in relatively thick tissue sections (e.g. 255m) and limited resolution along the Z axis during imaging, even when employing confocal microscopy. Array tomography circumvents these issues by antibody staining of ultrathin cryosections (e.g. 70nm) of nervous system tissue. There are numerous additional benefits to utilizing array tomography over traditional immunohistochemistry including the possibility of obtaining ultrastructural information from the tissue using scanning electron microscopy after immunofluorescence imaging and, perhaps most importantly for our purposes, the ability to perform multiple rounds of antibody staining of the same tissue section. As the position of the tissue is fixed on the slide, repeated staining with different antibodies against synaptic proteins will allow cataloging of which synaptic components are present or absent at all of the synapses in a single neuron. All five investigators in the user group have reached the limit of what can be achieved in assaying synapse morphology and function with traditional immunofluorescence methods. Thus, it is critical to furthering the research mission of these groups that the array tomography imaging technology be available on the Brandeis campus. PUBLIC HEALTH RELEVANCE: Numerous studies now point to defects in synapse formation as a possible cause for neurological disorders such as autism, mental retardation, and epilepsy. One approach to understanding how aberrant synapse formation contributes to these widespread neurological impairments is to first investigate how synapses are formed, maintained, and function in the non-pathological state using light microscopy. Acquisition of the Zeiss Axioimager Z2 System for Array Tomography will allow for unprecedented high-resolution imaging of synapses from the nervous system of a variety of experimental organisms.