Adaptive optics two photon microscopy with direct wavefront sensing using autofluorescent guide-stars

A fast direct wavefront sensing method for dynamic in-vivo adaptive optical two photon microscopy has demonstrated. By using the direct wavefront sensing and open loop control, the system provides high-speed wavefront measurement and correction. To measure the wavefront in the middle of a Drosophila embryo at early stages, autofluorescence from endogenous fluorophores in the yolk were used as reference guide-stars. This method does not rely on fluorescently labeled proteins as guide-stars, which can simplify the sample preparation for wavefront measurement. The method was tested through live imaging of a Drosophila embryo. The aberration in the middle of the embryo was measured directly for the first time. After correction, both contrast and signal intensity of the structure in the middle of the embryo was improved.