Grant: Research Tools and Instruments

With respect to the adage `seeing is believing', fluorescent microscopy has greatly advanced our understanding of cell biology by enabling researchers to visualize cells and the structures they contain. As cells are dynamic, live-cell imaging provides even more information. In this regard, if a picture is worth a thousand words, how much is a video worth? Recently, the advent of super-resolution imaging has enabled scientists zoom in even further into cells. However, there are trade-offs associated with many super-resolution approaches that limit their applicability for live-cell videos. Key among them is the fact that super-resolution imaging often requires higher laser power and longer exposure times, which can cause photobleaching (loss of signal), and phototoxicity (cell stress). Photobleaching limits the number of images that can be taken and the time-period over which imaging can occur. Phototoxicity means that you are not imaging cells under `normal' conditions, and can even lead to cell death. Additionally, longer exposure times limit the frame rate, restricting how often images can be captured, which means that you can miss dynamic cellular processes that occur quickly. Thus, researchers must compromise between image resolution, acquisition speed, and length of time imaging. This proposal is for upgrades to our current microscope system that will enable its use for live-cell super-resolution imaging by improving its capabilities and reducing laser exposure required for imaging. Specifically, a new camera and improved lenses will reduce the laser power and exposure time required to obtain images, placing less stress on the cells. Meanwhile, new hardware and software will automate the system, allowing it to operate faster, and provide additional imaging modalities. These upgrades will increase the flexibility of the system, enabling researchers to image cellular processes in real time that they have previously not been able to observe. Our team of investigators is interested in fundamental cell biological processes and shares a common need to image living cells at the highest resolution possible. We can achieve this goal by upgrading our existing SD-OSR microscope to the latest IXplore SpinSR system. The upgraded microscope is critical for enhancing the impact of each of our NSERC-funded programs, as this capability is not available elsewhere at the University of Calgary. Furthermore, the IXplore SpinSR system will be integrated into the Live Cell Imaging Laboratory at the University of Calgary, making it widely available to hundreds of researchers and further enhancing training capacity. These upgrades will unlock a whole new approach of imaging and will benefit Canada by facilitating a better understanding of many biological processes across many research fields.