Grant: Research Tools and Instruments

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.