Hypoxic workstation to conduct studies in low oxygen environments

在低氧（缺氧）条件下用细胞进行实验的挑战之一是在细胞处理后恢复到大气（21%）氧。细胞的变化可以发生在从低（缺氧或完全无氧（缺氧））到大气氧环境中去除细胞的5秒钟内。需要低氧条件的研究包括对细胞应激反应、炎症、癌症生物学、干细胞发育和细胞氧传感的研究。没有这些设备我们的工作是不可能的。目前，我们依靠三重气体（N2-O2-CO2）培养箱来产生缺氧条件。然而，这些孵化器现在已经有16年的历史了，已经超过了它们的预期寿命（10年），正在失败。由于我们每天都在使用它们，我们过时的三重气体孵化器被填满了。这些分庭的运作经常面临失败的威胁。在过去的一年里，关闭唐斯（更换O2和CO2传感器，水套泄漏，内部管道携带气体泄漏，控制主板故障）中断了Willmore和比格尔实验室的10名本科生，10名研究生和1名博士后实习生的实验。像这样的挫折浪费了宝贵的时间，并通过干扰学生的出版物，论文和完成本科论文项目所需的数据采集造成巨大的压力。事情已经到了临界点。如果在未来一年内没有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。实验室学员的项目依赖于这一基础设施，如果无法使用这一基础设施，他们的项目将受到极大的阻碍。所需的设备（InvivO 2 400）是一个缺氧工作站，配有封闭式手套箱、内置气体混合器和氧气传感器以及用于引入和取出细胞、培养基或组织培养消耗品的单独气锁。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缺氧条件。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，并随着时间的推移保持程序化的氧气条件（如分级低氧或预处理低氧暴露）。在卡尔顿大学没有类似的单位，在渥太华地区只有少数几个研究人员无法进入，用于细菌而不是哺乳动物细胞的研究。因此，在这方面，InvivO 2 400 Hypoprostimulus工作站将是该地区独一无二的。因此，低氧工作站对于Willmore和比格尔实验室以及合作实验室的研究计划的继续至关重要，这些研究计划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.