Single Molecule Tracking of Surface Glycans on Live Cells Using Metabolic Labeling and Click Chemistry
Albert Einstein College of Medicine
The surface of eukaryotic cells is covered with a dense layer of glycans that affect numerous physio- logical processes and aberrant cell-surface glycosylation has been implicated in cancer, in amma- tion and other diseases. Using a combination of bioorthogonal click reactions, super-resolution im- aging and single molecule tracking, we are able to observe the dynamics of cell-surface glycans on live cells with high spatial and temporal resolution. Our custom microscope also implements a form of confocal interference microscopy, based upon laser feedback interferometry, to simultaneously measure the topography of the membrane. We now seek to experimentally verify the predictions of our model which incorporates the energetics that govern the nucleation, growth and disassembly of early integrin adhesions in the presence of long glycan molecules. Our experimental approach has enabled the visualization of dynamic tunneling nanotubes connections between cells and the tracking of glycosylated receptors along the surface of these membranes.