Reduced Dyes Enhance Single-Molecule Localization Density for Live Superresolution Imaging
Cell-permeable rhodamine dyes are reductively quenched by NaBH4 into a non-fluorescent leuco-rhodamine form. Quenching is reversible, and their fluorescence is recovered when the dyes are oxidized. In living cells, oxidation occurs spontaneously, and can result in up to ten-fold higher densities of single molecule localizations, and more photons per localization as compared with unmodified dyes. These two parameters directly impact the achievable resolution, and we see a significant improvement in the quality of live-cell point-localization super-resolution images taken with reduced dyes. These improvements carry over to increase the density of trajectories for single-molecule tracking experiments.
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Carlini, L., Benke, A., Reymond, L., Lukinavicius, G., & Manley, S. (2014). Reduced Dyes Enhance Single-Molecule Localization Density for Live Superresolution Imaging. Chemphyschem, 15(4), 750–755. https://doi.org/10.1002/cphc.201301004
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WOS:000332747500025
2014
15
4
750
755
REVIEWED
EPFL