Prelabeling Expansion Single-Molecule Localization Microscopy with Minimal Linkage Error
| Autor: | Minsu Kang, Joo Yong Lee, Sangyoon Ko, Sang Hee Shim |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Streptavidin
Fluorescence-lifetime imaging microscopy Fluorophore 010405 organic chemistry Super-resolution microscopy Organic Chemistry Optical Imaging 010402 general chemistry 01 natural sciences Biochemistry Fluorescence Single Molecule Imaging 0104 chemical sciences chemistry.chemical_compound chemistry Microscopy Fluorescence Microscopy Biophysics Molecular Medicine Molecule Denaturation (biochemistry) Peptides Molecular Biology Fluorescent Dyes |
| Zdroj: | Chembiochem : a European journal of chemical biology. 22(8) |
| ISSN: | 1439-7633 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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