Abstract
Light microscopy has undergone something of a renaissance in cell biology since the advent of green-fluorescent protein (GFP) technology and the greater possibilities for imaging living cells. One of the key limitations of light microscopy is the resolution limit, known as the ‘diffraction resolution barrier’, resulting from the wave properties of light. A recent paper from the laboratory of Stefan Hell in Göttingen, Germany, describes a technique facilitating an increase in the optical resolution obtained by light microscopy (Fig. 1) 1. Klar T.A. et al. Proc. Natl. Acad. Sci. USA. 2000; 97: 8206-8210 Crossref PubMed Scopus (1360) Google Scholar . The process, termed point spread function engineering by stimulated emission depletion (PSFE by STED), results in an increase in optical resolution from the 250–300 nm range classically observed with low-numerical-aperture oil-immersion objectives to 90–100 nm. Confocal sectioning is improved approximately five fold. Clearly, this type of technology will have many invaluable applications in the coming years.
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