Exploring Membrane Microdomain Organization by Electron Microscopy

The flow of information through cells requires the constant remodeling of cell signaling and trafficking networks. To observe the remodeling events associated with receptor activation, we have generated and analyzed high resolution topographical maps of colloidal gold nanoprobes marking receptors and signaling proteins in native membranes. The technology involves labeling membrane sheets stripped from the dorsal cell surface with functionalized nanoprobes, imaging the labeled sheets by TEM, extracting probe coordinates from digitized images and analyzing the distributions of the probes with respect to each other and to membrane features like clathrincoated pits and caveolae.