Single-Molecule Image Scanning Microscopy

In image scanning microscopy (ISM), a small array detector is placed in the image plane of a confocal microscope. For each scan position of the focused excitation beam, the array detector images the emitted fluorescence signal. This combination of structured illumination and structured detection improves the spatial resolution of the laser-scanning microscope by a factor of two compared to conventional confocal microscopy. While ISM will replace conventional confocal laser-scanning microscopy [1], the combination of structured illumination and structured detection in an ISM scheme for single-molecule imaging has not yet been explored.