Fluorescence photobleaching/photoactivation imaging system upgrade

Fluorescent-labeling of biological molecules has become a powerful tool for the analysis of cellular events using the light microscope. Several quantitative microscopy techniques that use fluorescent labels have been developed that allow the study of proteins, nucleic acids and lipids within living cells. Fluorescence Recovery After Photobleaching (FRAP) and Photoactivation (PA), are two techniques used to measure the diffusion of biological molecules and to track their movement. Our 5 research groups are requesting FRAP/PA Unit to upgrade a state-of-the-art spinning-disk confocal laser microscopy system. The FRAP/PA Unit provides the ability to accurately localise and track the rapid movement of biological molecules in 3 dimensions (3D), with limited loss of fluorescence over time, and with reduced cellular damage. As such, this upgrade significantly increases the capabilities of the existing infrastructure. The FRAP/PA Unit will support the research programs of our 5 groups, which encompass a range of fundamental studies in basic science, including: the study of how cells respond to DNA damage; how nerve cells in the brain develop; how lipids regulate cellular processes; how proteins such as collagen form complex structures; and how the biological machinery that drives smooth muscle contraction functions. Once installed, this upgrade module will make the CFI funded microscope a one of kind instrument in Atlantic Canada, providing a unique training opportunity for students to learn advanced quantitative light microscopy techniques. The requested equipment will also be accessible to research groups other than those named in this application thereby facilitating the research of many other investigators at Dalhousie University.