PT  - JOURNAL ARTICLE
AU  - Christoph Spahn
AU  - Jonathan B. Grimm
AU  - Luke D. Lavis
AU  - Marko Lampe
AU  - Mike Heilemann
TI  - Bleaching-independent, whole-cell, 3D and multi-color STED imaging with exchangeable fluorophores
AID  - 10.1101/420638
DP  - 2018 Jan 01
TA  - bioRxiv
PG  - 420638
4099  - http://biorxiv.org/content/early/2018/09/18/420638.short
4100  - http://biorxiv.org/content/early/2018/09/18/420638.full
AB  - We demonstrate bleaching-independent STED microscopy 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 in fixed and living cells. Foremost, 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, multi-color and live cell STED microscopy with up to 100 min acquisition time.