Adaptive optics has been used to compensate the detrimental effects of aberrations in a range of high-resolution microscopes. We investigate how backscattered laser illumination can be used as the source for direct wavefront sensing using a pinhole-filtered Shack-Hartmann wavefront sensor. It is found that the sensor produces linear response to input aberrations for a given specimen. The gradient of this response is dependent upon experimental configuration and specimen structure. Cross sensitivity between modes is also observed. The double pass nature of the microscope system leads in general to lower sensitivity to odd-symmetry aberration modes. The results show that there is potential for use of this type of wavefront sensing in microscopes.

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