LIVE-PAINT allows super-resolution microscopy inside living cells using reversible peptide-protein interactions

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 liveS. 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. Oi et al. describe LIVE-PAINT, a new method to achieve superresolution fluorescent imaging within live cells. They demonstrate its efficacy by imaging a number of different proteins inside live S. cerevisiae cells. Their method is widely applicable, easily implemented with extended data acquisition times.