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

摘要: With continuing advances in the resolving power of super-resolution microscopy, inefficient labeling proteins with suitable fluorophores becomes a limiting factor. For example, low density achieved antibodies or small molecule tags limits attempts to reveal local protein nano-architecture cellular compartments. On other hand, high laser intensities cause photobleaching within and nearby an imaged region, thereby further reducing impairing multi-plane whole-cell 3D imaging. Here, we show that both can be addressed by repetitive application trisNTA-fluorophore conjugates reversibly binding histidine-tagged novel approach called single-epitope imaging (SERI). single-plane demonstrate that, after multiple rounds imaging, signal is increased. Using same washing re-labeling, compensated for above below plane. This proof-of-principle study demonstrates provides versatile solution break 'labeling barrier' bypass multi-plane, experiments.