Purchase a Beckman Coulter MoFlo Astrios Flow Cytometer

描述（由申请人提供）：这是一项收购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），增加容量并增加当前细胞分选设施的能力，这是NIH资助的10个主要研究项目的关键组成部分。提出这一请求的理由是根据以下因素进行的认真评估：1.需要提高流式细胞仪的细胞分选服务能力，这对目前NIH资助的研究人员和Wistar研究所项目的预期增长至关重要。 2. 2种分拣机之一的FACSAria II能够进行16种荧光染料/4种激光检测，而现有的MoFlo仅允许5种荧光染料。升级将使调查人员从多色分析转向分类时具有更大的灵活性和效率，而不是将他们限制在一台分类机内。 3.需要生物安全防护等级高于BSL-1的实验目前仅限于FACSAria II，造成时间安排上的瓶颈。所提出的新细胞分选仪将适当配备，以允许BSL-2+分选。 4.由于缺乏适当的激光激发，荧光蛋白分选目前是有限的。所提出的分选机将具有绿色/黄色范围内的激光，用于最佳激发荧光蛋白，例如mCherry、mTomato等。所提出的分选机将能够进行六向分选，从而提供分离比当前四个种群更多种群的能力，沿着能够分选到多达1536个威尔斯孔的载玻片或平板中。 6.现有的MoFlo Legacy仍然能够很好地执行，并且几乎每天都在使用，并且由于其速度和样品回收而受到许多人的青睐，但是由于与年龄相关的维修，停机时间越来越频繁。这导致操作员时间的增加，并将NIH资助的项目置于风险之中，因为只有一台分拣机可能会导致更长的停机时间。虽然大修和改进是一种选择，但所涉费用最好用于新文书。这个备受推崇的设施，成立于30多年前，目前由三名经验丰富的技术人员组成，并由教师用户委员会监督。Wistar和邻近机构的许多研究人员经常使用细胞分选服务，由于如上所述的容量限制，现有的分选机经常被大量预订，偶尔会出现积压。因此，我们建议通过购买一台设备齐全的MoFlo Astrios高速分选机来升级分选能力，以保持对10个主要用户和8个次要用户的最先进支持，其中包括41个R 01/P01/U 01 NIH资助的研究奖项，该分选机具有适当的生物安全水平遏制、六向分选以及将单细胞直接分选到组织培养板或载玻片中的能力。

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.