Shack-Hartmann wavefront-sensor-based adaptive optics system for multiphoton microscopy.
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.
Cha
Jae Won
JW
Massachusetts Institute of Technology, Department of Mechanical Engineering, Cambridge, Massachusetts 02139, USA.
Ballesta
Jerome
J
So
Peter T C
PT
D000818:Animals; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005246:Feedback; D007089:Image Enhancement / Q000295:instrumentation*; D016247:Information Storage and Retrieval / Q000379:methods*; D007909:Lenses; D051379:Mice; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014159:Transducers
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