There is an unequivocal need to develop and employ advanced techniques in microscopy toward understanding the cellular functions of glycans. Glycans participate in many cellular properties, including cell structure, cell-cell recognition, protein trafficking, and protein-protein interaction. Hence, changes in glycan abundance or composition can have considerable effects on cellular function, and as a consequence human health. Advanced imaging techniques are becoming a vital part of an integrative approach to understanding the roles of glycans in health and disease. These techniques provide insights into molecular composition, while preserving the spatial context of the chemical, structural, and physiological responses to changes in glycans. Recent advances in identify lectin specificities and the availability of a broad range of fluorescently labeled and biotinylated lectins has rapidly advanced our ability to identify changes in the location and abundance of specific glycans in cells and tissues. Moreover, several powerful approaches in confocal microscopy, including FRET, FRAP, and FLIP, permit highly sensitive measures of molecular dynamics within living cells. These measures will help researchers understand the fundamental effect glycans have on cellular function. The central goal of this proposal is to bring the ability to employ advanced imaging techniques to the GlyCORE investigators in their diverse research programs and to help develop new approaches in the cell biology of glycans. These goals will be accomplished in three aims: 1) establish an imaging core that contains three different imaging platforms, including a powerful new confocal microscope; 2) support the GlyCORE investigators in their individual projects by working to develop new imaging applications; and 3) develop an extensive training program to develop expertise and awareness of new methods in imaging and image analysis. This proposal will transform research in the glycosciences at the University of Mississippi by providing access to advanced instrumentation and expertise and training in the use of these imaging systems.
