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J Biomed Opt. 2010 Jul-Aug;15(4):046022. doi: 10.1117/1.3475954.

Shack-Hartmann wavefront-sensor-based adaptive optics system for multiphoton microscopy.

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1
Massachusetts Institute of Technology, Department of Mechanical Engineering, Cambridge, Massachusetts 02139, USA.

Abstract

The imaging depth of two-photon excitation fluorescence microscopy is partly limited by the inhomogeneity of the refractive index in biological specimens. This inhomogeneity results in a distortion of the wavefront of the excitation light. This wavefront distortion results in image resolution degradation and lower signal level. Using an adaptive optics system consisting of a Shack-Hartmann wavefront sensor and a deformable mirror, wavefront distortion can be measured and corrected. With adaptive optics compensation, we demonstrate that the resolution and signal level can be better preserved at greater imaging depth in a variety of ex-vivo tissue specimens including mouse tongue muscle, heart muscle, and brain. However, for these highly scattering tissues, we find signal degradation due to scattering to be a more dominant factor than aberration.

PMID:
20799824
PMCID:
PMC2937046
DOI:
10.1117/1.3475954
[Indexed for MEDLINE]
Free PMC Article
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