Pupil-segmentation-based adaptive optics for microscopy

Na Ji; Daniel E. Milkie; Eric Betzig

doi:10.1117/12.876398

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14 February 2011 Pupil-segmentation-based adaptive optics for microscopy

Na Ji, Daniel E. Milkie, Eric Betzig

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Proceedings Volume 7931, MEMS Adaptive Optics V; 79310I (2011) https://doi.org/10.1117/12.876398
Event: SPIE MOEMS-MEMS, 2011, San Francisco, California, United States

Abstract

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, the direct wavefront-sensing approach commonly used in astronomy is not suitable for most biological samples due to their strong scattering of light. 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, local tilt in the wavefront is measured from image shift. The aberrated wavefront is then obtained either by measuring the local phase directly using interference or with phase reconstruction algorithms similar to those used in astronomical AO. 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.

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Na Ji, Daniel E. Milkie, and Eric Betzig "Pupil-segmentation-based adaptive optics for microscopy", Proc. SPIE 7931, MEMS Adaptive Optics V, 79310I (14 February 2011); https://doi.org/10.1117/12.876398

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