Purchase a Beckman Coulter MoFlo Astrios Flow Cytometer

描述(由申请人提供)：这是一份购买MoFlo Astrios流式细胞仪/高速细胞分选仪的提案，以取代旧的MoFlo Legacy(2001)，增加现有细胞分选设施的能力，这是美国国立卫生研究院资助的10个主要研究项目的关键组成部分。提出这一要求的理由基于基于以下因素的仔细评估：1.需要改善流式细胞术设施中的细胞分选服务能力，这对目前由NIH资助的研究人员和Wistar Institute项目的预期增长至关重要。2.FACSAria II是两台分选机中的一台，它能够进行16次荧光铬/4激光探测，而现有的MoFlo只能探测5次荧光。升级将使调查人员从多颜色分析转向分类时具有更大的灵活性和效率，而不是仅限于一种分类器。3.要求生物安全遏制水平高于BSL-1的试验目前仅限于FACSAria II，导致进度瓶颈。拟议的新细胞分选机将配备适当的设备，以便进行BSL-2+分选。4.由于缺乏适当的激光激发，目前荧光蛋白质分选受到限制。拟议的分选机将具有绿/黄色范围内的激光，以最佳地激发荧光蛋白质，如mCherry、mTomato等。5.拟议的分选机将能够进行六路分选，提供比目前四个分选机更多的种群的能力，以及能够分选到多达1536个孔的玻片或平板中。6.现有的MoFlo Legacy仍然能够运行良好，基本上每天都在使用，由于其速度和样品回收而受到许多人的青睐，但由于与年龄有关的维修，停机时间越来越频繁和越来越长。这导致操作员的时间增加，并将NIH资助的项目置于风险之中，因为只有一个分拣机可能导致更长的停机时间。虽然大修和改进是一种选择，但所涉及的费用将更好地应用于新工具。这个备受赞誉的机构成立于30多年前，目前有三名经验丰富的技术人员，并由一个教师用户委员会进行监督。Wistar和邻近机构的许多调查人员经常使用细胞分拣服务，由于上文讨论的容量限制，现有的分拣服务经常被大量预订，偶尔会有积压。因此，我们建议升级分选能力，以保持对10个主要用户和8个次要用户的最先进支持，获得41个R01/P01/U01 NIH资助的研究奖项，方法是购买设备齐全的MoFlo Astrios高速分选机，具有适当的生物安全级别遏制、六向分选，并能够将单个细胞直接分选到组织培养板或玻片中。

DESCRIPTION (provided by applicant): This is a proposal to acquire a MoFlo Astrios flow cytometer/high-speed cell sorter to replace an old MoFlo Legacy (2001), increasing capacity and adding to capabilities of the current cell sorting facility, a key component for 10 major NIH-funded research programs. The rationale for this request rests on a careful assessment based on the following factors: 1. A need to improve cell sorting service capacity in the Flow Cytometry Facility as crucial for the current NIH-funded investigators and the anticipated growth in Wistar Institute programs. 2. One of 2 sorters, the FACSAria II, is capable of 16 fluorochrome/4 laser detection while the existing MoFlo allows for only five fluorochromes. Upgrading would allow greater flexibility and efficiency for investigators moving from multi-color analysis to sorting, not restricting them to only one sorter. 3. Experiments requiring bio-safety containment at levels greater than BSL-1 are currently restricted to the FACSAria II, resulting in scheduling bottlenecks. The proposed new cell sorter would be appropriately equipped to allow for BSL-2+ sorting. 4. Fluorescent protein sorting is currently limited, given the lack of appropriate laser excitation. The proposed sorter would have a laser in the green/yellow range for optimal excitation of fluorescent proteins such as mCherry, mTomato, etc. 5. The proposed sorter would be capable of six-way sorting, providing the ability to separate more populations than the current four, along with the ability to sort into slides or plates up to 1536 wells. 6. Te existing MoFlo Legacy is still capable of performing well and used on a mostly daily basis, and is preferred by many for its speed and sample recovery, but is suffering from more frequent and increasing amounts of downtime due to age-related repairs. This results in increasing amounts of operator time and places NIH-funded projects at risk due to the potential for longer downtimes due to availability of only one sorter. While major overhaul and enhancement is an option, the expense involved would be better applied into a new instrument. This well-regarded facility, established over 30 years ago, is currently staffed by three experienced technicians and monitored by a faculty user committee. Many investigators at Wistar and neighboring institutions routinely use the cell sorting services, with the existing sorters often heavily booked with occasional backlogs due to capacity limitations as discussed above. Therefore, we propose to upgrade sorting capacity to maintain state- of-the-art support for 10 major and 8 minor users, with 41 R01/P01/U01 NIH-funded research awards, through purchase of a well-equipped MoFlo Astrios high-speed sorter, with appropriate bio-safety level containment, six-way sorting, and the ability to sort single cells directly into tissue culture plates or slides.