Maintaining and Enhancing Flow Cytometry Capacity for Biological Research at the University of Guelph

所需设备说明：本申请用于购买一台21色流式细胞仪。我们使用流式细胞术来快速同时识别来自动物血液或组织的数千个不同的细胞。这些可能是来自动物的细胞，我们正在测试一种新的疫苗，需要看看它的免疫细胞或T细胞有什么反应。细胞被放入流式细胞仪，在那里它们会通过激光，而激光会对它们进行“电击”。特定的细胞类型以特定的方式“破坏”激光束，这使我们能够对复杂混合物中的所有细胞组进行分类。而且，像T细胞这样对疫苗有反应的细胞，其表面的蛋白质与没有反应的细胞不同。我们可以在这些蛋白质上贴上“细胞标记”，就像名字标签一样。这些姓名标签被激光照射时会发出荧光。我们知道T细胞是否对疫苗有反应，因为没有名字标签的细胞(没有反应的细胞)通过激光时不会发出荧光。在医学和科学中，能够用尽可能多的名称标签分析复杂的细胞组是很重要的--21色流式细胞术是一种非常强大的工具，可以让我们做到这一点。它可以让我们用更少的动物细胞来分析单个样本中的更多细胞，因此我们可以更快地得出诊断或重要的科学结论。有8名尖端科学家和30名学生，加上来自奎尔夫大学的其他研究人员，他们使用我们目前的流式细胞仪，但它只有8色的容量，而且经常出现故障。因此，我们正在申请资金购买最先进的21色流式细胞仪来更换我们的旧机器。这是Cytekr北极光多色流式细胞仪，将被安置在病理生物学系的“核心设施”中。该设施有专门的技术和研究人员来维护机器，并帮助满足我们NSERC资助的研究人员对机器的许多不同用途。这种尖端的流式细胞仪将提高我们对学生的研究和培训的质量，这将使我们能够在全球范围内竞争，但也将造福于加拿大的动物和人类。支持的工作性质：这种流式细胞仪是一种基本的研究工具，用于识别小鼠、猪和绵羊的疫苗特异性T细胞反应，用于表征鸟类、猫、狗、马和牛的癌症和改变的免疫细胞，用于了解是什么导致哮喘或细菌感染失控，以及肿瘤如何劫持身体以创造自己的血液供应。对加拿大的预期结果和益处：这项研究对于开发基于免疫的干预措施和病毒载体疫苗、改进几种癌症的诊断、测试癌症和哮喘的新疗法以及确定细菌感染的免疫控制至关重要，这对加拿大的动物和人类都有好处。

Description of equipment requested: This application is for the purchase of a 21-colour flow cytometry machine. We use flow cytometry to rapidly and simultaneously identify thousands of different cells coming from an animal's blood or tissue. These could be cells from an animal in which we are testing a new vaccine and need to see how its immune cells, or T cells, have responded. Cells are put into the flow cytometer where they go past lasers which "zap" them. Specific cell types "break" the laser beam in specific ways, which allows us to classify all the cell groups in a complex mixture. But also, cells like T cells that have responded to a vaccine have different proteins on them than cells that have not responded. We can attach "cell markers" to those proteins that are like name tags. These name tags fluoresce when they are zapped by the laser. We know whether the T cells have responded to the vaccine because cells without the name tag (the ones that didn't respond) go past the laser without fluorescing. In medicine and science, it is important to be able to analyze complex groups of cells with as many name tags as possible - 21-colour flow cytometry is a very powerful tool that allows us to do that. It lets us analyze more cells from a single sample, using fewer cells from the animal, so we arrive at a diagnosis or an important scientific conclusion much more quickly. There are 8 cutting-edge scientists and >30 students, plus other researchers from across the University of Guelph that use our current flow cytometer, but it has only 8-colour capacity and breaks down frequently. So, we are requesting funds to purchase a state-of-the-art 21-colour flow cytometer to replace our old machine. It is the Cytekr Northern Lights multicolour flow cytometer, and will be housed in the Department of Pathobiology's "core facility". This facility has dedicated technical and research staff to maintain the machine and to help meet the many different uses that our NSERC-funded researchers have for it. This cutting-edge flow cytometer will increase the quality of our research and training of our students, which will allow us to compete globally, but will also benefit both the animals and people of Canada. Nature of work to be supported: This flow cytometer is an essential research tool for identifying vaccine-specific T cell lymphocyte responses in mice, pigs and sheep, for characterizing cancerous and altered immune cells from birds, cats, dogs, horses and cattle, for understanding what causes asthma or loss of control of bacterial infections, and how tumours hijack the body to create their own blood supply. Anticipated outcomes and benefits to Canada: This research is key to developing immune-based interventions and viral vectored vaccines, improving diagnosis of several types of cancer, testing novel therapies for cancer and asthma, and for defining immune control of bacterial infections, which benefits both the animals and people of Canada.