High-Content Super-Resolution Imaging of Live Cell by uPAINT

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.