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Whole-Cell, 3D, and Multicolor STED Imaging with Exchangeable Fluorophores
Christoph Spahn1, Jonathan B Grimm2, Luke D Lavis2
1Institute of Physical and Theoretical Chemistry , Goethe-University Frankfurt , Max-von-Laue-Str. 7 , 60438 Frankfurt , Germany.
Abstract:
We demonstrate stimulated emission depletion (STED) microscopy of whole bacterial and eukaryotic cells using fluorogenic labels that reversibly bind to their target structure. A constant exchange of labels guarantees the removal of photobleached fluorophores and their replacement by intact fluorophores, thereby circumventing bleaching-related limitations of STED super-resolution imaging. We achieve a constant labeling density and demonstrate a fluorescence signal for long and theoretically unlimited acquisition times. Using this concept, we demonstrate whole-cell, 3D, multicolor, and live-cell STED microscopy.
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