Journal
FEBS LETTERS
Volume 573, Issue 1-3, Pages 6-10Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.febslet.2004.07.035
Keywords
near-field scanning optical microscopy; single molecule detection; sub-diffraction limit; cell surface distribution; microdomain; DC-SIGN
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Clustering of cell surface receptors into microdomains in the plasma membrane is an important mechanism for regulating cellular functions. Unfortunately, these domains are often too small to be resolved with conventional optical microscopy. Near-field scanning optical microscopy (NSOM) is a relatively new technique that combines ultra high optical resolution, down to 70 nm, with single molecule detection sensitivity. As such, the technique holds great potential for direct visualisation of domains at the cell surface. Yet, NSOM operation under liquid conditions is far from trivial. In this contribution, we show that the performance of NSOM can be extended to measurements in liquid environments using a diving bell concept. For the first time, individual fluorescent molecules on the membrane of cells in solution are imaged with a spatial resolution of 90 nm. Furthermore, using this technique we have been able to directly visualise nanometric sized domains of the C-type lectin DC-SIGN on the membrane of dendritic cells, both in air and in liquid. (C) 2004 Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.
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