Adaptive optics for fluorescence wide-field microscopy using spectrally independent guide star and markers

We describe the implementation and use of an adaptive optics loop in the imaging path of a commercial wide field microscope. We show that it is possible to maintain the optical performances of the original microscope when imaging through aberrant biological samples. The sources used for illuminating the adaptive optics loop are spectrally independent, in excitation and emission, from the sample, so they do not appear in the final image, and their use does not contribute to the sample bleaching. Results are compared with equivalent images obtained with an identical microscope devoid of adaptive optics system.