Adaptive optics (AO) is necessary to correct aberrations when imaging the mouse eye with high numerical aperture. In order to obtain cellular resolution, we have implemented wavefront sensorless adaptive optics for in vivo fluorescence imaging of mouse retina. Our approach includes a lens-based system and MEMS deformable mirror for aberration correction. The AO system was constructed with a reflectance channel for structural images and fluorescence channel for functional images. The structural imaging was used in real-time for navigation on the retina using landmarks such as blood vessels. We have also implemented a tunable liquid lens to select the retinal layer of interest at which to perform the optimization. At the desired location on the mouse retina, the optimization algorithm used the fluorescence image data to drive a modal hill-climbing algorithm using an intensity or sharpness image quality metric. The optimization requires ~30 seconds to complete a search up to the 20th Zernike mode. In this report, we have demonstrated the AO performance for high-resolution images of the capillaries in a fluorescence angiography. We have also made progress on an approach to AO with pupil segmentation as a possible sensorless technique suitable for small animal retinal imaging. Pupil segmentation AO was implemented on the same ophthalmic system and imaging performance was demonstrated on fluorescent beads with induced aberrations.
Daniel J. Wahl, Stefano Bonora, Oscar S. Mata, Bengt K. Haunerland, Robert J. Zawadzki, Marinko V. Sarunic, and Yifan Jian, "Wavefront sensorless approaches to adaptive optics for in vivo fluorescence imaging of mouse retina," Proc. SPIE 9717, Adaptive Optics and Wavefront Control for Biological Systems II, 97170A (Presented at SPIE BiOS: February 13, 2016; Published: 15 March 2016); https://doi.org/10.1117/12.2213572.
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