Cytometry analyzer capable of fast fluorescence-activated cell sorting

背景研究环境微生物的高通量高效方法对于了解感兴趣物种的多样性，丰度和生态学至关重要。例如，有必要监测受气候变化影响的地区或有毒蓝细菌（例如，加拿大大西洋沿岸地区，湖泊和河流），以确定影响微生物物种的存在和丰度的环境因素，这些微生物物种与基本生态过程有关，如初级生产（光合作用活动），营养物质循环和负责生产影响人类活动的有毒化合物。我们急需一台现代化的细胞分选仪。我在新玩法大学（UNB）的研究小组使用流式细胞术（FC）和荧光激活细胞分选的组合研究了感兴趣的环境微生物，这使我们能够1）在短时间内仔细检查数百个微生物细胞（即，秒至分钟）和2）分离具有感兴趣特征的细胞（例如，大小、光合特性）。FC和FACS的灵敏度和速度在细胞分选仪（CS）中是商业上可获得的，其从根本上超过了仅基于光学显微镜和视觉识别的微生物学研究的效率。在过去的11年里，我的研究小组一直受益于使用CS，但由于实际过时（低分拣速度），该仪器已经达到了其使用寿命的终点，更重要的是，鉴于我们拥有的型号已经停产，服务合同将于明年暂停，维护维修成本不断增加。这两个因素迫使我们使用私人流式细胞仪设施，以保证获得最先进的CS，并继续保持我们的研究贡献的高质量。雇用CS服务是一个短期的绝望的解决方案，以增加我们的研究目标的范围和影响，但从长远来看，这是不切实际的，因为增加每个实验的成本严重限制了我们可以做的研究数量。我们迫切需要一个高性能的CS，以降低我们的研究成本，并恢复我们的实验灵活性。所要求的CS重新统一了分析能力和分选效率，保证了我们当前和未来研究目标的持续。依靠第三方细胞仪设备延迟了我们的研究目标和NSERC支持的论文项目（5 HQP）。此外，新的CS将有利于来自四个不同的UNB研究小组的20名HQP的培训。生物学系和UNB副总裁研究致力于支持我们机构的微生物研究，并为此申请提供70，000美元的现金。此外，我还从供应商那里获得了63 340美元的实物捐助。这两项捐款的总和占估计总费用的51%以上。

Background. High-throughput efficiency methods to study environmental microbes are essential to understand the diversity, abundance and ecology of species of interest. For example, it is necessary to monitor the composition of algal and bacterial communities in areas affected by climate change, or with recurrent proliferation of toxic cyanobacteria (e.g., coastal regions, lakes, and rivers in Atlantic Canada), to identify the environmental factors affecting the presence and abundance of microbial species associated to fundamental ecological processes such as primary production (photosynthetic activity), nutrient recycling and responsible for the production of toxic compounds that impact human activities. Our urgent need of a modern cell sorter. My research group at the University of New Brunswick (UNB) studies environmental microbes of interest using a combination of flow cytometry (FC) and fluorescence-activated cell sorting, which give us the capacity to 1) scrutinize hundreds of microbial cells in a short time (i.e., second to minutes) and 2) isolate cells with characteristics of interest (e.g., size, photosynthetic properties). The sensitivity and speed of FC and FACS are commercially available in cell sorters (CS), which surpass radically the efficiency of microbiology studies based solely on light microscopy and visual identification. In the last eleven years, my research group has had the benefit of using a CS, but that instrument has reached the end of its useful life due to practical obsolescence (low sorting speed) and, importantly, the increasing maintenance-repair cost, given that the model we own has been discontinued and service contracts will be suspended next year. Those two factors have forced us to use private flow cytometry facilities to guarantee access to state-of-the-art CS and to continue and maintain the high quality of our research contributions. Hiring CS services is a short-term desperate solution to increase the reach and impact of our research aims, but it is impractical in the long term given that the increased cost per experiment limits severely the number of studies we can do. We have an urgent need for a high-performance CS to reduce the cost of our research and to recover our experimental flexibility. The requested CS reunites the analytical capabilities and sorting efficiency that guarantee the continuation of our current and future research aims. Relaying on a third-party cytometric facility is delaying our research objectives and thesis projects (5 HQP) supported by NSERC. Additionally, the new CS will benefit the training of 20 HQP from four different UNB research groups. The Department of Biology and UNB Vice President Research are committed to supporting microbial research in our institution, and are contributing $70,000 in cash to this application. Also, I have secured a $63,340 in-kind contribution from the vendor. The sum of both contributions represents >51% of the estimated total cost.