Leica AOBS SP2 confocal for the MCB/DBS Imaging Facility

适用于 MCB/DBS 成像设备的 Leica AOBS SP2 共焦

• 批准号: 6732286
• 负责人: Jodi M. Nunnari
• 金额: $ 49.64万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6732286
• 关键词: bioimaging /biomedical imaging; biomedical equipment purchase; confocal scanning microscopy; lasers

DESCRIPTION (provided by applicant): We propose to purchase a Leica TCS SP2 AOBS laser scanning confocal microscope equipped with special 405 nm diode and 594 nm orange HeNe lasers to be housed in our existing light microscopy Imaging Facility. This facility was used by 77 different researchers in calendar year 2002, including 56 using our fiveyear- old Leica SP1 UV confocal. The growth and increasing sophistication of our users have revealed two significant shortcomings of the existing microscopes. First is the inability to perform key experiments, including 1) fluorescence recovery after photobleaching (FRAP), 2) photo-activation of fluorophores in defined regions of interest, and 3) time-resolved imaging of more than one component at rates faster than one two-component image every 1-2 seconds. Second is a debilitating lack of sensitivity from our existing SP1 confocal for even moderately bright samples. This leads many experiments better suited for a confocal to be done on our deconvolution microscope, despite the optical flaws, simply to get adequate sensitivity. A straightforward way of addressing all of these concerns is the replacement of our existing confocal with one of contemporary design. Of contemporary confocals, the Leica TCS SP2 AOBS and Zeiss LSM 510 META are best suited to meet the needs of our most sophisticated users while still allowing a multi-user facility. We have selected the Leica TCS SP2 AOBS because 1) the Leica can meet our experimental needs and has superior sensitivity as evidenced during demos by Leica and Zeiss; 2) the superior Leica "filter-free" optical design; 3) the continuity of user interface for our large user group, making extensive retraining unnecessary; 4) continuity of our strong relationship with our Leica field service engineer, and 5) the trade in value of our existing Leica SP1. Major NIH-funded users will be Jodi M. Nunnari ("Regulation of Mitochondrial Fission"), Jonathon M. Scholey ("Roles of Microtubule-Based Motility in Mitosis and Intracellular Transport"), and Carol A. Erickson ("Direct Observation of Neural Crest Cell Emigration in a Living Embryo"). These users will account for approximately 50% of the instrument time, with the balance being offered to our large imaging community. This use will continue to be under the direction of one PhD and one MS scientist and administered under the current policies of an existing departmental Imaging Committee. A letter showing strong institutional support for maintenance and continued operation of the confocal is included.

描述（由申请人提供）：我们建议购买Leica TCS SP2 AOBS激光扫描共聚焦显微镜，配备特殊的405 nm二极管和594 nm橙子HeNe激光器，安装在我们现有的光学显微镜成像设备中。2002年，77名不同的研究人员使用了该设备，其中56人使用了我们已有5年历史的Leica SP1紫外共焦设备。我们用户的增长和日益复杂揭示了现有显微镜的两个显著缺点。首先是无法进行关键实验，包括1）光漂白后的荧光恢复（FRAP），2）在限定的感兴趣区域中的荧光团的光活化，以及3）以比每1-2秒一个双组分图像更快的速率对多于一个组分进行时间分辨成像。第二个是我们现有的SP1共焦系统对于中等亮度的样品缺乏灵敏度。这导致许多实验更适合在我们的去卷积显微镜上进行共焦，尽管有光学缺陷，只是为了获得足够的灵敏度。解决所有这些问题的一个简单方法是用当代设计取代我们现有的共焦。 在当代共焦镜中，Leica TCS SP2 AOBS和Zeiss LSM 510 Meta最适合满足我们最复杂的用户的需求，同时仍然允许多用户设施。我们选择Leica TCS SP2 AOBS是因为：1）Leica能够满足我们的实验需求，并具有上级灵敏度，这在Leica和Zeiss的演示中得到了证明; 2）Leica的上级“无滤镜”光学设计; 3）用户界面的连续性适合我们庞大的用户群，无需进行大量的再培训; 4）我们与徕卡现场服务工程师的牢固关系的延续性，以及5）我们现有徕卡SP1的价值交易。 NIH资助的主要用户将是Jodi M。Nunnari（“线粒体分裂的调节”），Jonesman M. Scholey（“有丝分裂和细胞内运输中基于微管的运动性的作用”）和Carol A. Erickson（“Direct Observation of Neural Crest Cell Emigration in a Living Embryo”）.这些用户将占仪器使用时间的大约50%，其余部分将提供给我们的大型成像社区。这种使用将继续在一名博士和一名MS科学家的指导下进行，并根据现有部门成像委员会的现行政策进行管理。一封信显示出强大的机构支持的维护和继续运作的共聚焦。