Near-field microscopy of living cells

While optical microscopy is an everyday tool in the biological sciences, significant advances could be made if optical resolution were increased. We have evaluated the utility of near-field optical microscopy, with its demonstrated sub-diffraction optical resolution, for fluorescence studies of living cells. We show that the incorporation of a fluorescent feedback method, photon- density feedback, permits near-field studies of living cells. With this approach it is possible to make local high- resolution point measurements from living cells without impacting the physiology of the system under study. By integrating near-field illumination fibers, with confocal detection, traditional cameras and PMT-based detection methods, we have developed a near-field microscope for biological studies which also permits traditional lens-based optical studies of biological system With this system we are able to make immediate comparisons between confocal and near-field data from the same biological system.

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