Acquisition of a DeltaVision OMX SR 3D SIM system for super-resolution imaging

描述（由申请人提供）：本申请申请资金用于购买Applied Precision DeltaVision OMX SR 3D结构化照明系统（3D- sim），安装在纽约洛克菲勒大学的生物成像资源中心（BIRC）。DeltaVision OMX SR （DV OMX SR）是第一个商业上可用的“超分辨率”显微镜之一，允许研究人员在所有三个轴上以两倍的分辨率解决生物成分或结构，这可以使用任何传统显微镜实现。此外，这种改进的分辨率可以在10-20微米厚的任何类型的样品中同时实现多种颜色，使其成为一种非常灵活的方法，现在可以应用于解决迄今为止无法回答的生物医学研究中的许多重要问题。最初受益于该系统的研究人员将在广泛的生物体和健康相关问题上工作，包括：端粒长度在抑制哺乳动物细胞DNA损伤中的作用（与癌症和年龄相关的疾病有关）；哺乳动物细胞分裂过程中确保复制染色体精确分离的分子基础（评估化学成分抑制剂作为癌症治疗药物）；在学习和疾病（阿尔茨海默氏症、精神分裂症、帕金森症）期间发生的神经元树突棘结构变化；了解链球菌和其他病原体的结构；了解酵母和哺乳动物核孔复合体蛋白的组织和动力学（也涉及细胞分裂过程中适当的染色体分离，并且已知是有效的肿瘤抑制因子）；以及阐明真核生物细胞核中DNA复制位点的排序（遗传物质的准确复制对于基因组稳定性和避免诸如癌症之类的体细胞遗传疾病至关重要）。所有这些问题都将通过使用荧光标记来定位固定细胞内蛋白质、分子或大分子结构的相对位置的方法来解决。虽然这种方法已经被应用，但在所有情况下，即使是BIRC中的高端显微镜也无法区分相邻组件，这些组件在xy轴上的距离小于200nm，或者在z轴上的距离小于500nm，这严重限制了我们可以提出的问题。我们在这里表明，使用DV OMX SR系统的初步实验在我们所有的研究领域提供了新的生物学信息，开辟了新的科学探究线，如下所示。所要求的系统将被安置在位于纽约中心位置的一个大型且支持良好的核心设施中，该设施可以访问众多外部研究人员以及指定用户和其他RU调查人员。

DESCRIPTION (provided by applicant): This application requests funds to purchase an Applied Precision DeltaVision OMX SR 3D Structured Illumination system (3D-SIM) to be installed in the Bio-Imaging Resource Center (BIRC) at The Rockefeller University in New York. The DeltaVision OMX SR (DV OMX SR) is one of the first commercially available "super-resolution" microscopes that permit's researchers to resolve biological components or structures at double the resolution, in all three axes, of that which can be achieved using any conventional microscope. Moreover, this improved resolution can be achieved with multiple colors simultaneously and in any type of sample up to 10-20 micrometers thick, rendering this a very flexible approach that can now be applied to address a number of important questions in biomedical research that were hitherto impossible to answer. The researchers who will initially benefit from this system work on a broad array of organisms and health-related issues, including: the role of telomere length in repressing DNA damage in mammalian cells (relevant to cancer and age-related diseases); the molecular basis for ensuring accurate segregation of replicated chromosomes during mammalian cell division (evaluating chemical inhibitors of components as drugs for cancer therapy); structural changes in the dendritic spines of neurons that occur during learning as well as in disease (Alzheimer's, schizophrenia, Parkinsonism); understanding the structure of Streptococcal bacteria and other pathogens; understanding the organization and dynamics of both yeast and mammalian nuclear pore complex proteins (also implicated in proper chromosome segregation during cell division, and known to be potent tumor suppressors); and elucidating the ordering of DNA replication sites in the eukaryotic nucleus (accurate copying of the genetic material being crucial for genomic stability and to avoid somatic genetic diseases such as cancer). All of these questions will be approached by the method of using fluorescent tags to locate the relative positions of proteins, molecules or macromolecular structures within fixed cells. Although this approach has already been applied, in all cases, the inability of even the high-end microscopes in the BIRC to distinguish between neighboring components that are closer together than 200 nm in the xy axis, or 500 nm in the z-axis, has severely limited the questions we can ask. We show here that preliminary experiments using the DV OMX SR system provided new biological information in all of our research areas, opening up new lines of scientific enquiry that are presented below. The requested system will be housed in a large and well-supported core facility in a central location in New York, which offers access to numerous outside researchers as well as the named users and other RU investigators.