1. LIVE-PAINT: Super-Resolution Microscopy Inside Live Cells Using Reversible Peptide-Protein Interactions
- Author
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Simon G. J. Mochrie, Louise Holyoake, Owen Kantelberg, Curran Oi, Mathew H. Horrocks, Lynne Regan, and Zoe Gidden
- Subjects
chemistry.chemical_classification ,0303 health sciences ,Fluorophore ,Super-resolution microscopy ,Oligonucleotide ,Biomolecule ,Peptide ,02 engineering and technology ,021001 nanoscience & nanotechnology ,Fluorescence ,Protein–protein interaction ,03 medical and health sciences ,chemistry.chemical_compound ,chemistry ,Biophysics ,0210 nano-technology ,Peptide sequence ,030304 developmental biology - Abstract
We present LIVE-PAINT, a new approach to super-resolution fluorescent imaging inside live cells. In LIVE-PAINT only a short peptide sequence is fused to the protein being studied, unlike conventional super-resolution methods, which rely on directly fusing the biomolecule of interest to a large fluorescent protein, organic fluorophore, or oligonucleotide. LIVE-PAINT works by observing the blinking of localized fluorescence as this peptide is reversibly bound by a protein that is fused to a fluorescent protein. We have demonstrated the effectiveness of LIVE-PAINT by imaging a number of different proteins inside live S. cerevisiae. Not only is LIVE-PAINT widely applicable, easily implemented, and the modifications minimally perturbing, but we also anticipate it will extend data acquisition times compared to those previously possible with methods that involve direct fusion to a fluorescent protein.
- Published
- 2020
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