The biggest obstacle for deep tissue imaging is the scattering of light due to the heterogeneous distribution of biological tissue. In this respect, multiphoton microscopy has an inherent advantage as the scattering is significantly reduced by the use of longer excitation wavelengths. However, as we go deeper into the brain, effects of scattering still accumulate resulting in a loss of resolution and increased background noise. Adaptive optics is an ideal tool of choice to correct for such distortions of the excitation wavefront; the incident light can be tuned to cancel out the wavefront distortion experienced while propagating into greater depths resulting in a diffraction limited focus at the depth of interest.
However, the biggest limitation of adaptive optics for in vivo brain imaging is its limited corrected field-of-view (FOV). For typical multiphoton laser scanning microscopes, the wavefront corrector for adaptive optics is placed at the pupil plane. This means that a single correction wavefront is applied to the entire scanned FOV which results in inefficient correction as the correction is averaged over the entire FOV. In this work, we demonstrate a novel approach to measure and display different correction wavefronts over different segments of the FOV. The application of the different correction wavefronts for each segment is realized in parallel resulting in fast aberration corrected imaging over a large FOV for high resolution in vivo brain imaging.
Jung-Hoon Park and Meng Cui, "Large field-of-view wavefront control for deep brain imaging
(Conference Presentation)," Proc. SPIE 9717, Adaptive Optics and Wavefront Control for Biological Systems II, 97170E (Presented at SPIE BiOS: February 13, 2016; Published: 27 April 2016); https://doi.org/10.1117/12.2213566.4848767665001.
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