Hypoxic workstation to conduct studies in low oxygen environments

在低氧(低氧)条件下进行细胞实验的挑战之一是在细胞处理后返回到大气(21%)氧。细胞从低氧(低氧或完全无氧(缺氧))环境移至大气氧气环境后5分钟内可发生细胞变化。需要低氧条件的研究包括细胞应激反应、炎症、癌症生物学、干细胞发育和细胞氧气感应的研究。没有这台设备，我们的工作是不可能的。目前我们依靠三重气体(N_2-O_2-CO_2)孵化器来产生低氧条件。然而，这些孵化器现在已经16岁了，正在失败，已经超过了它们的预期寿命(即10年)。由于我们每天都在使用它们，我们过时的三重气体孵化器已经满了。这些商会在持续不断的失败威胁下运作。在过去的一年里，停机维修(更换O2和CO2传感器、水套泄漏、内部输气管道泄漏、主板控制故障)扰乱了威尔莫尔和比格实验室的10名本科生、10名研究生和1名博士后的实验。像这样的挫折浪费了宝贵的时间，并造成了巨大的压力，因为干扰了学生为出版、论文和完成本科论文项目所需的数据获取。一切都快到临界点了。如果明年不能用更好的系统取代这些系统，那么我们的核心研究计划(依赖于低氧环境的产生)将无法继续下去。依赖于此基础设施的实验室的学员的项目将因无法使用而受到极大阻碍。*所需的设备(InvivO2 400)是一个低氧工作站，带有一个封闭的手套箱、一个内置的气体混合器和氧气传感器以及一个单独的气闸，用于引入和移除细胞、培养液或组织培养消耗品。拟议的仪器将允许在细胞操作期间控制环境，并确保在整个实验过程中保持低氧条件。低氧工作站将使我们的实验室(以及合作实验室)能够在细胞操作和收获时保持低氧条件，并随着时间的推移编程氧气条件(如分级低氧或预适应低氧暴露)。卡尔顿大学没有类似的单位，渥太华地区只有几个单位是研究人员无法接触到的，用于细菌而不是哺乳动物细胞的研究。因此，InvivO2 400低氧工作站在这方面是该区域独一无二的。因此，低氧工作站对于威尔莫尔和比格实验室以及合作实验室的研究计划的继续至关重要，这些实验室依赖于低氧环境的产生。

One of the challenges of experimentation with cells under low oxygen (hypoxic) conditions is the return to atmospheric (21%) oxygen after cell treatment. Cellular changes can occur WITHIN 5 MINUTES of removal of cells from low (hypoxic or complete absence of oxygen (anoxic)) to atmospheric oxygen environments. Research requiring hypoxic conditions includes studies on cell stress response, inflammation, cancer biology, stem cell development and oxygen sensing by cells. Our work is impossible without this equipment. Currently we are relying upon triple gas (N2-O2-CO2) incubators to generate hypoxic conditions. However, these incubators are now 16 years old and are failing, having reached beyond their expected lifespan (which is 10 years). As we use them on a daily basis, our out-dated triple gas incubators are filled to capacity. These chambers operate under constant threat of failure. Over the past year, shut downs for repairs (replacement of O2 and CO2 sensors, leakage of the water jacket, leakage in internal tubing carrying gases, failure of controlling motherboards) has disrupted the experiments of 10 Undergraduate Students, 10 Graduate Students and 1 Postdoctoral trainees in the Willmore and Biggar labs. Set-backs like this waste precious time and cause enormous stress by interfering with data acquisition by students needed for publications, theses, and completion of undergraduate thesis projects. Things are reaching a breaking point. If these are not replaced with a better system within the next year, then our core research program (which relies on the generation of hypoxic environments) will not be able to continue. The projects of the trainees of the labs dependent upon this infrastructure will be greatly hindered by not having access to it.***The equipment requested (InvivO2 400) is a hypoxic workstation with an enclosed glove box, a built-in gas mixer and oxygen sensor and a separate airlock for introduction and removal of cells, media or tissue culture consumables. The proposed instrumentation would allow for controlled environments during cell manipulation and ensure that hypoxic conditions are maintained throughout the experiment. The hypoxic workstation will enable our lab (as well as collaborating labs) to maintain low oxygen conditions upon cell manipulation and harvest, as well as have programmed oxygen conditions (such as graded hypoxia or preconditioning hypoxia exposures) over time. There are no similar units at Carleton University and only a few within the Ottawa region that are inaccessible to the researchers and used for studies with bacteria and not mammalian cells. The InvivO2 400 Hypoxic Workstation would therefore be unique to the region in this respect. The hypoxic workstation is therefore CRITICAL for the continuation of the research programs of the Willmore and Biggar labs, as well as collaborating labs, that are dependent upon the generation of low oxygen environments.