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Adaptive optics have been used to compensate the detrimental effects of aberrations in a range of high resolution microscopes. Aberration measurement has been implemented in various way, using direct wave front sensing or indirect optimisation methods. We investigate how backscattered laser illumination can be used as the source for direct wave front sensing using a pinhole filtered Shack Hartmann wave front 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. The double pass nature of the microscope system leads to lower sensitivity to odd-symmetry aberration modes. © 2012 SPIE.

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