Acquisition of a Zeiss LSM 900 confocal microscope with Airyscan 2 for an Imaging and Microscopy Core

研究综述这项拨款提案寻求为蔡司LSM 900激光扫描共焦提供资金 配备AiryScan 2的显微镜，适用于使用高级成像和显微镜核心的NIH资助研究人员 洛玛琳达大学的设施，进行组织化学分析和细胞和 纸巾。一方面，该系统将用于利用荧光对各种蛋白质和细胞进行定位 标记方法，而另一方面，它将被用来检查活细胞和组织的活动。 这两种成像技术是必需的，因为NIH资助的主要和次要用户群体有不同的 需要。他们的研究涵盖心血管生理学、怀孕、发育、神经生物学、 神经内分泌学、癌症生物学、放射医学、再生医学和健康差距，但都是 在测量细胞之间的空间位置，蛋白质表达的变化， 蛋白质在细胞内的定位，以及与细胞信号有关的空间和时间序列。这个 用户组的研究记录很好，代表了一些最成功的调查人员 机构。蔡司LSM 900共焦将支持许多NIH研究拨款和调查 校园计划和培训计划。核心设施有两台共用的共焦显微镜 接近或在其功能使用结束时。收购带有AiryScan2的蔡司LSM 900将解决这一问题 紧迫的问题，并将使资助的研究人员能够继续他们的研究，包括细胞信号转导，原位 杂交、蛋白质表达和定位、细胞和组织形态、分子间相互作用 和蛋白质、空间重建、时空关系和亚细胞运输。这些问题 需要能够对固定或活体制剂中的单个或多个荧光探针进行成像。基于 根据资助项目的需要，蔡司LSM 900共焦显微镜是一种理想的替代仪器。 该仪器有两个高灵敏度共焦通道用于记录荧光；一个用于透射光 (DIC)，并具有用于超分辨率成像的AiryScan 2单元以及自适应对焦功能，并且可以 执行马赛克平铺以进行多维重建。Airyscan2还将允许快速共焦 细胞信号系统的空间和时间记录的成像，这是多个 奖项。蔡司LSM 900提供了一个完全集成的系统，可用于广泛的研究 并将提供与现有多光子蔡司710系统的连续性。该机构承认 设施和用户群体的质量，十多年来一直全力支持该设施 为人员配备、供应和服务合同提供资金。PI在共焦方面有相当丰富的经验 显微镜和将监督仪器的日常操作，并成为多学科内部的一部分 监督核心设施的咨询委员会。该仪器将安置在现有的AIM设施中， 为这个使用率很高的共用成像核心提供持续的现代仪器。

Research Summary This grant proposal seeks funding for a Zeiss LSM 900 laser scanning confocal microscope with Airyscan 2 for NIH funded investigators who use the Advanced Imaging and Microscopy Core Facility at Loma Linda University to perform both histochemical analyses and real-time imaging of cells and tissues. On the one hand, the system would be used to localize various proteins and cells using fluorescent labeling approaches, while on the other it would be used to examine the activity of living cells and tissues. These two imaging technologies are required as the NIH funded major and minor user group has diverse needs. Their research encompasses cardiovascular physiology, pregnancy, development, neurobiology, neuroendocrinology, cancer biology, radiation medicine, regenerative medicine, and health disparities, but all have common interests in measuring the spatial apposition between cells, changes in protein expression, localization of proteins within cells, and spatial and temporal sequences involved with cell signaling. The research record of the user group is excellent, representing some of the most successful investigators at the institution. The Zeiss LSM 900 confocal would support numerous NIH research grants and investigative programs across campus and training programs. The core facility has two shared-use confocal microscopes that are near or at their end of functional use. Acquisition of a Zeiss LSM 900 with a AiryScan 2 will solve this pressing issue and will enable funded researchers to continue their studies including cell signaling, in-situ hybridization, protein expression and localization, cell and tissue morphology, interactions between molecules and proteins, spatial reconstruction, spatial-temporal relationships, and subcellular transport. These questions require the ability to image single or multiple fluorescent probes in either fixed or live preparations. Based on the needs of the funded projects, a Zeiss LSM 900 confocal microscope is an ideal replacement instrument. The instrument has 2 high-sensitivity confocal channels for recording fluorescence; one for transmitted light (DIC) and has an Airyscan 2 unit for super-resolution imaging along with adaptive focusing capabilities and can perform mosaic tiling for multidimensional reconstruction. The Airyscan 2 will also allow for rapid confocal imaging for spatial and temporal recordings of cell signaling systems, which are key aspects to multiple awards. The Zeiss LSM 900 offers a fully integrated system available for the wide breadth of studies of the user group and will provide continuity with an existing multiphoton Zeiss 710 system. The institution recognizes the quality of the facility and user group and has strongly supported the facility for over a decade with full funding for staffing, supplies, and service contracts. The PI has considerable experience with confocal microscopy and will oversee the daily operation of the instrument and be part of a multidisciplinary internal advisory committee that oversees the core facility. The instrument will be housed in the existing AIM facility, providing continued contemporary instrumentation for this highly used shared-use imaging core.