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High-resolution light microscope reveals the fundamental mechanisms of nerve communication

In a simultaneous publication (Science Express, 13th April 2006), STED microscopy revealed the spatial distribution of the bruchpilot protein and aided neurobiologists from the European Neuroscience Institute and the University of Würzburgin understanding the protein's central role in the formation of active synaptic zones. STED microscopy radically distinguishes itself from conventional farfield light microscopy in the fact that its resolution is no longer fundamentally limited by the wavelength of light used. Using STED, nanoscale optical studies are now possible inside cells.

Since its discovery in the 17th century, the light microscope has been the key to new biological and medical discoveries. Light, however, propagating as a wave, is subject to the phenomenon of diffraction, whose resolution-limiting effects were first described by Ernst Abbe in 1873. Abbe observed that structures which were closer to each other than ~200nm could not be visually separated when observed using visible light; when viewed through the optical microscope they are perceived as a blurred, single entity. Abbe's realization of the resolution limitation of the optical microscope was long thought to be a unalterable law of far-fieldlight imaging. Achieving higher resolution required the use of an electron microscope.

Despite that fact that electron beams can be more tightly focused, it is often difficult to efficiently label the proteins of a cell to render them visible with an electron microscope. Moreover, electron beams are only able to penetrate the first several micrometers of a biological sample. For these reasons, among others, despite using electron microscopy for high-resolution cell imaging, many questions of nerve function remained unanswered. In contrast, using fluorescent molecules as markers, one can specifically label individual proteins with high efficiency, rendering them visible with the conventional fluorescent microscope. Unfortunately the high resolutio
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Source:Max-Planck-Gesellschaft


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