Abstract
In this chapter, we present the uPAINT method (Universal Point Accumulation Imaging in Nanoscale Topography), a simple single-molecule super-resolution method which can be implemented on any wide field fluorescence microscope operating in oblique illumination. The key feature of uPAINT lies in recording high numbers of single molecules at the surface of a cell by constantly labeling while imaging. In addition to generating super-resolved images, uPAINT can provide dynamical information on a single live cell with large statistics revealing localization-specific diffusion properties of membrane biomolecules. Interestingly, any membrane biomolecule that can be labeled with a fluorescent ligand can be studied, making uPAINT an extremely versatile method.
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Acknowledgments
We wish to thank B. Lounis for helpful discussions. This research was funded by Centre National de la Recherche Scientifique (CNRS), the Région Aquitaine and the Agence Nationale pour la Recherche (ANR), the Fondation pour la Recherche Médicale, and the European Union’s seventh framework program for research and development ERC grant Nano-Dyn-Syn.
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Giannone, G., Hosy, E., Sibarita, JB., Choquet, D., Cognet, L. (2013). High-Content Super-Resolution Imaging of Live Cell by uPAINT. In: Sousa, A., Kruhlak, M. (eds) Nanoimaging. Methods in Molecular Biology, vol 950. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-137-0_7
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DOI: https://doi.org/10.1007/978-1-62703-137-0_7
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