Adaptive optics two-photon scanning laser fluorescence microscopy

Yaopeng Zhou; Thomas Bifano; Charles Lin

doi:10.1117/12.875596

14 February 2011 Adaptive optics two-photon scanning laser fluorescence microscopy

Yaopeng Zhou, Thomas Bifano, Charles Lin

Proceedings Volume 7931, MEMS Adaptive Optics V; 79310H (2011) https://doi.org/10.1117/12.875596
Event: SPIE MOEMS-MEMS, 2011, San Francisco, California, United States

Abstract

Two-photon fluorescence microscopy provides a powerful tool for deep tissue imaging. However, optical aberrations from illumination beam path limit imaging depth and resolution. Adaptive Optics (AO) is found to be useful to compensate for optical aberrations and improve image resolution and contrast from two-photon excitation. We have developed an AO system relying on a MEMS Deformable Mirror (DM) to compensate the optical aberrations in a two-photon scanning laser fluorescence microscope. The AO system utilized a Zernike polynomial based stochastic parallel gradient descent (SPGD) algorithm to optimize the DM shape for wavefront correction. The developed microscope is applied for subsurface imaging of mouse bone marrow. It was demonstrated that AO allows 80% increase in fluorescence signal intensity from bone cavities 145um below the surface. The AO-enhanced microscope provides cellular level images of mouse bone marrow at depths exceeding those achievable without AO.

Citation Download Citation

Yaopeng Zhou, Thomas Bifano, and Charles Lin "Adaptive optics two-photon scanning laser fluorescence microscopy", Proc. SPIE 7931, MEMS Adaptive Optics V, 79310H (14 February 2011); https://doi.org/10.1117/12.875596

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