Acquisition of a New Confocal Microscope to Replace an Obsolete Instrument

描述(由申请人提供)：1993年，W.M.凯克基金会向威斯康星大学麦迪逊分校授予了第一笔赠款，以建立生物成像实验室(www.keck.BioImaging.witc.edu)。该实验室购买了一台Bio-Rad MRC1024共焦显微镜，并立即开始为来自密歇根大学校园的研究人员提供机会，将共焦成像纳入他们的研究中。11年后，在NCRR的奖励下，该实验室增加了一台Bio-Rad Radiance 2100 MP共焦/多光子显微镜，使开放源代码的多光子显微镜首次可在华盛顿大学麦迪逊分校使用。仅在过去的12个月里，威斯康星大学麦迪逊分校的近60个实验室使用了该实验室的显微镜，这些实验室代表着生物科学的大约20个广泛研究领域。现在，该实验室希望用一台新的显微镜取代原来的Bio-Rad显微镜。虽然MRC-1024已经是一种有能力的仪器15年了，但现在它几乎每月都需要维修，而卡尔·蔡司决定在去年9月之后不再为MRC-1024提供服务合同，这让这个问题变得更加复杂。此外，按照当代标准，MRC-1024已经过时。这反映在过去12个月中，随着实验室用户将他们的工作转移到Radiance 2100 MP，MRC-1024的使用量减少了60%以上。不幸的是，Radiance无法容纳所有希望使用它的人，这给一些用户造成了严重的延误。此外，许多用户操作Radiance 2100 MP仅用于共焦成像，从而影响了显微镜作为多光子仪器的主要作用。在选择MRC-1024的替代品时，对徕卡公司(TCS SP5)、尼康(Eclipse C1Plus)、蔡司(LSM 510 Meta)和奥林巴斯(FluoView FV1000)的显微镜进行了评估，以确定哪种仪器最适合在多用户成像实验室中操作。在比较了提到的每个系统，直接与每个系统一起工作，并查看了它们的可靠性和服务记录后，我们得出的结论是，奥林巴斯FluoView FV1000在为研究人员提供各种研究项目和经验的实验室中操作时，提供了灵活性、易用性和性能之间的最佳平衡。要求的FV1000将配备六条激光线路、两个标准光电倍增管采集通道和两个光谱探测器。光谱探测器将使用户能够分解光谱重叠的信号，这是减少自发荧光造成的干扰的强大工具。分离光谱重叠的信号还将在设计实验时提供更大的灵活性，例如，在同一实验中使用EGFP和EYFP，或在同时成像多个荧光团时。公共卫生相关性： 这项申请申请资金购买一个新的共聚焦显微镜，将由八名生物医学研究科学家使用。其中七名科学家将使用显微镜研究与癌症、贫血和心脏老化等各种疾病有关的过程。剩下的科学家将利用显微镜研究胚胎大脑中不断生长的轴突如何进行适当的连接，以确保正常的大脑发育。

DESCRIPTION (provided by applicant): In 1993, the W.M. Keck Foundation awarded the first grant to UW-Madison to establish a Laboratory for Biological Imaging (www.keck.bioimaging.wisc.edu). The Laboratory acquired a Bio-Rad MRC 1024 confocal microscope, and began immediately to offer investigators from across the UW campus the opportunity to incorporate confocal imaging in their research. Eleven years later, with an award from NCRR, the Laboratory added a Bio-Rad Radiance 2100 MP confocal/multiphoton microscope, making open source multiphoton microscopy available for the first time at the UW-Madison. In the last 12 months alone, nearly 60 laboratories at UW-Madison, representing approximately 20 broad research areas in the biological sciences, have used the Laboratory's microscopes. Now the Laboratory wishes to replace its original Bio-Rad microscope with a new microscope. While the MRC-1024 has been a capable instrument for 15 years, it now needs repairs almost monthly, and compounding this problem is Carl Zeiss's decision not to offer a service contract for the MRC- 1024 after September of last year. In addition, the MRC-1024 is obsolete by contemporary standards. This is reflected by a reduction of over 60% in usage of the MRC-1024 during the past 12 months, as the Laboratory's users shift their work to the Radiance 2100 MP. Unfortunately, the Radiance cannot accommodate all of those who wish to use it, creating significant delays for some users. In addition, many users operate the Radiance 2100 MP solely for confocal imaging, compromising the microscope's primary role as a multiphoton instrument. In selecting a replacement for the MRC-1024, microscopes from Leica Microsystems (TCS SP5), Nikon (Eclipse C1Plus), Zeiss (LSM 510 META ) and Olympus (FluoView FV1000) were evaluated to determine which instrument would be the best suited for operation in a multi-user imaging laboratory. After comparing each of the systems mentioned, working directly with each, and reviewing their reliability and service records, we have concluded that the Olympus FluoView FV1000 offers the best balance of flexibility, ease of use, and performance for operating in a Laboratory that serves investigators with a diverse range of research projects and experience. The FV1000 requested will be equipped with six laser lines, two standard PMT collection channels, and two spectral detectors. The spectral detectors will enable users to unmix spectrally overlapping signals, a powerful tool for reducing interference by caused by autofluorescence. Separating spectrally overlapped signals also will provide increased flexibility in designing experiments e.g., using eGFP and eYFP in the same experiment, or in imaging simultaneously multiple fluorophores. PUBLIC HEALTH RELEVANCE: This application request funds to purchase a new confocal microscope that will be used by eight biomedical research scientists. Seven of these scientists will use the microscope to study processes that are involved in various diseases such as cancer, anemia, and cardiac aging. The remaining scientist will employ the microscope to study how growing axons in the embryonic brain make proper connections to ensure normal brain development.