Enhanced labeling density and whole-cell 3D dSTORM imaging by repetitive labeling of target proteins.

ABSTRACT: With continuing advances in the resolving power of super-resolution microscopy, the inefficient labeling of proteins with suitable fluorophores becomes a limiting factor. For example, the low labeling density achieved with antibodies or small molecule tags limits attempts to reveal local protein nano-architecture of cellular compartments. On the other hand, high laser intensities cause photobleaching within and nearby an imaged region, thereby further reducing labeling density and impairing multi-plane whole-cell 3D super-resolution imaging. Here, we show that both labeling density and photobleaching can be addressed by repetitive application of trisNTA-fluorophore conjugates reversibly binding to a histidine-tagged protein by a novel approach called single-epitope repetitive imaging (SERI

SUBMITTER: Venkataramani V 

PROVIDER: S-EPMC5882651 | biostudies-literature | 2018 Apr

REPOSITORIES: biostudies-literature