Abstract
Near-field optical microscopy (NSOM), combined with the contrast techniques, particularly fluorescence, of conventional, far-field microscopy, promises to yield high-resolution images of the molecular organisation of cell membranes. We have previously resolved structures in lipid monolayers, and in the membranes of cells after fixation and drying. Both of these preparations are relatively flat and it is not difficult to maintain the scanning tip in near-field proximity to the surface. Wet samples, particularly live cells, are much more difficult objects than dried samples. Control of scanning is complicated by the damping effects of the liquid above the cells and by the roughness and softness of cell surfaces. We have constructed a NSOM designed to maintain living cells in buffer and will describe the ways in which the problems raised can be overcome. We have been able to image both fixed and live cells in liquid and have used fluorescent antibodies, lectins, and lipid analogues as probes of membrane organization in these samples.
© 1997 Optical Society of America
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