We present a novel system for adaptive optics two photon imaging. We utilize the bandwidth of the femtosecond excitation beam to perform coherence gated imaging (OCT) of the sample. The location of the focus is directly observable in the cross sectional OCT images, and adjusted to the desired depth plane. Next, using real time volumetric OCT, we perform Wavefront Sensorless Adaptive Optics (WSAO) aberration correction using a multi-element adaptive lens capable of correcting up to 4th order Zernike polynomials. The aberration correction is performed based on an image quality metric, for example intensity. The optimization time is limited only by the OCT acquisition rate, and takes ~30s. Following aberration correction, two photon fluorescence images are acquired, and compared to results without adaptive optics correction. This technique is promising for multiphoton imaging in multi-layered, scattering samples such as eye and brain, in which traditional wavefront sensing and guide-star sensorless adaptive optics approaches may not be suitable.
Yifan Jian, Michelle Cua, Stefano Bonora, Edward N. Pugh, Robert J. Zawadzki, and Marinko V. Sarunic, "Coherence gated wavefront sensorless adaptive optics for two photon excited fluorescence retinal imaging
(Conference Presentation)," Proc. SPIE 9712, Multiphoton Microscopy in the Biomedical Sciences XVI, 97121E (Presented at SPIE BiOS: February 16, 2016; Published: 27 April 2016); https://doi.org/10.1117/12.2213655.4848767250001.
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