Spahn Christoph, Grimm Jonathan B, Lavis Luke D, Lampe Marko, Heilemann Mike
Institute of Physical and Theoretical Chemistry , Goethe-University Frankfurt , Max-von-Laue-Str. 7 , 60438 Frankfurt , Germany.
Janelia Research Campus , Howard Hughes Medical Institute , 19700 Helix Drive , Ashburn , Virginia 20147 , United States.
Nano Lett. 2019 Jan 9;19(1):500-505. doi: 10.1021/acs.nanolett.8b04385. Epub 2018 Dec 14.
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.
我们展示了使用与目标结构可逆结合的荧光标记对完整细菌和真核细胞进行受激发射损耗（STED）显微镜成像。标记的持续交换保证了光漂白荧光团的去除以及它们被完整荧光团的替代，从而规避了STED超分辨率成像中与漂白相关的限制。我们实现了恒定的标记密度，并证明在长时间以及理论上无限制的采集时间内都能获得荧光信号。利用这一概念，我们展示了全细胞、三维、多色和活细胞STED显微镜成像。
