Whole-Cell, 3D, and Multicolor STED Imaging with Exchangeable Fluorophores.

Spahn, Christoph; Grimm, Jonathan B; Lavis, Luke D; Lampe, Marko; Heilemann, Mike · Nano Lett · 2019

basic_science · Level V

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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.