Inhomogeneous optical properties of biological samples make it difficult to obtain diffraction-limited resolution in depth. Correcting the sample-induced optical aberrations needs adaptive optics (AO). However,  due to their strong scattering of light, the direct wavefront-sensing approach commonly used in astronomy is not suitable for most biological samples. We developed an image-based AO approach that is insensitive to sample scattering. By comparing images of the sample taken with different segments of the pupil illuminated, the aberrated wavefront is measured from image shift. We implemented this pupil-segmentation-based approach in a two-photon fluorescence microscope and demonstrated that diffraction-limited resolution can be recovered from nonbiological and biological samples.

Pupil-segmentation-based adaptive optics for microscopy

242nd National Meeting (2011)

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