In this thesis we investigated the ability of two-photon multifocal microscopy for single-molecule microscopy in live cells and organisms. Two-photon excitation combined with multifocal scanning... Show moreIn this thesis we investigated the ability of two-photon multifocal microscopy for single-molecule microscopy in live cells and organisms. Two-photon excitation combined with multifocal scanning has the potential to achieve, high (temporal) resolution imaging at a low background. Splitting the laser beam into multiple beamlets reduced laser power to all but eliminate photobleaching. The low background combined with the fast scan speeds and absence of photobleaching allowed us to measure single-particles and single-molecules in live zebrafish embryo's for long time periods. To the best of our knowledge, it is the first time that single-molecule molecules have been observed in a wide-field two-photon microscope. Show less
