Regensburg 2004 – scientific programme

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SYLS: Life Sciences on the Nanometer Scale - Physics Meets Biology

SYLS 3: Symposium "Life Sciences on the Nanometer Scale - Physics Meets Biology"

SYLS 3.10: Poster

Wednesday, March 10, 2004, 16:00–18:30, B

High-Resolution Near-Field Fluorescence Microscopy of Single Nuclear Pore Complexes in an Intact Nuclear Envelope — •C. Höppener¹, J.-P. Siebrasse², U. Kubitscheck², R. Peters², H. Fuchs¹, and A. Naber³ — ¹Physikalisches Institut, Westfälische Wilhelms-Universität, 48149 Münster — ²Institut für Medizinische Physik und Biophysik, Westfälische Wilhelms-Universität, 48149 Münster — ³Institut für Angewandte Physik, Universität Karlsruhe (TH), 76131 Karlsruhe

The nuclear pore complex (NPC) is a large macromolecular protein assembly embedded in the nuclear envelope (NE) of a eukaryotic cell and mediates the highly selective, bi-directional exchange of all kinds of molecules between cytoplasm and nucleus. So far single NPCs could not be resolved by optical means since the NPCs are densely packed in the membrane. We present high-resolution near-field optical images of a fluorescently labelled NE [1] performed in a buffer solution. These images represent the first example of a high-resolution scanning near-field optical measurement of a functionally intact biomembrane. A super-resolution of 60nm enables us to optically identify for the first time single, closely neighboured NPCs. Furthermore, the appearance of the NPCs markedly varies with the primary antibody used for fluorescence labelling, thus revealing information about the location of a specific nucleoporin within the NPC.

[1] C. Höppener, D. Molenda, H. Fuchs, and A. Naber, J. Microsc. 210, 288 (2003)