Precision localization of cellular proteins with fluorescent Fab-based probes

Abstract

With continuously improving resolution of today’s (super-resolution) microscopes, a major technical limitation of light microscopy based image analysis is linkage error – a visualization error that is measured by the distance between the cellular target to be detected and the fluorescence emitter used for detection. The linkage error of standard labelled antibodies is caused by the size of the antibody and the random distribution of fluorescent emitters on the antibody surface. In this study, we describe a class of staining reagents that effectively reduce the linkage error by more than five-fold when compared to conventional staining techniques. We believe this class of reagents realize an unmet need in cell biological super resolution imaging studies where the precise localization of the target of interest is crucial for the understanding of complex biological phenomena