Grant: Research Tools and Instruments - Category 1 (<$150,000)

My research utilizes electrophysiology, cell imaging, and cell culturing to investigate the effects of oxygen availability on the behaviour of stem cells. For example, in the brain, cells exist in oxygen (O2) environments that range from 2% O2 to 10% O2. However, in the research setting, cells are often subjected to 95% air (21% O2). Thus it is apparent that the experimental O2 levels used and O2 values experienced by cells in the body are inconsistent. Different levels of O2 have the capability of modifying protein function and gene expression and thus can affect stem cell behaviour. Alterations in protein function can lead to entry of calcium, which in turn can initiate regulation of genes involved in stem cell growth and development into different cell types. Since O2 levels have the capacity to modify cellular activity (e.g. protein function and gene regulation) it is important to determine how these cells typically behave in the body. Equipment use: The equipment will be used to study the interaction between exposure of stem cells to physiologically relevant oxygen 'physioxia' and stem cell differentiation. The Biospherix chamber will be required to maintain specific O2 levels when growing cells and tissue. The basis of cellular function and communication is via membrane electrical potentials. The amplifier and digidata are used to detect the electrical activity of cells. The micromanipulators are necessary to accurately place electrodes on the cells (fine control). As any vibrations can significantly affect the ability to put electrodes onto cells, vibrations will be isolated via an anti-vibration table. The DG-4 will be utilized to measure intracellular calcium levels. Using DG-4's capability for rapid switching between light filters (<1 msec) in addition to use of Fura-2AM (a dye that binds calcium and fluoresces when bound), measurements of intracellular calcium can be made. The CCD camera will be used to transmit the images generated with the DG-4 to the computer for analysis. This camera can also be used for deep tissue scanning and training of students.