| Affiliation: | 1. Center for Molecular Spectroscopy and Dynamics, Institute for Basic Science (IBS), Seoul, 02841 Republic of Korea
Department of Chemistry, Korea University, Seoul, 02841 Republic of Korea;2. Department of Biomedical Engineering, Ulsan National Institute of Science and Technology(UNIST), Ulsan, 44919 Republic of Korea;3. Center for Molecular Spectroscopy and Dynamics, Institute for Basic Science (IBS), Seoul, 02841 Republic of Korea |
| Abstract: | Expansion microscopy combined with single-molecule localization microscopy (ExSMLM) has a potential for approaching molecular resolution. However, ExSMLM faces multiple challenges such as loss of fluorophores and proteins during polymerization, digestion or denaturation, and an increase in linkage error arising from the distance between the fluorophore and the target molecule. Here, we introduce a trifunctional streptavidin to link the target, fluorophore and gel matrix via a biotinylizable peptide tag. The resultant ExSMLM images of vimentin filaments demonstrated high labeling efficiency and a minimal linkage error of ～5 nm. Our ExSMLM provides a simple and practical means for fluorescence imaging with molecular resolution. |
