The application of adaptive optics to microscopy is important for the correction of the aberrations due to the mismatch of refractive index of the sample and the imperfections present in the optics used. Adaptive optics in microscopy does not require elevated operating frequencies however, inserting a wavefront sensor and an adaptive element (e.g. a deformable mirror) within the microscope optics, is not straightforward and brings to complicated optical setup. Past works have used wavefront sensorless techniques based on the process of image sharpening . Our work instead, focused on designing a new and simple alternative solution that can be easily implemented in the microscope setup with a very compact device. Pupil segmentation is a recent wavefront sensing technique, based on the division of the pupil in smaller sub-apertures. In these sub-pupils, the wavefront is locally comparable to a flat tilted wavefront, with a different slope in each aperture. These tilt variations are evaluated directly from the imaging sensor (i.e. a CCD camera) from the displacement of the PSF spots of the sup-pupils from a reference spot. It is in this way possible to calculate the average gradient and, from an appropriate matrix calculation, reconstruct the wavefront.
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