Enhancing image quality in cleared tissue with adaptive optics

Area of Science:

• Biomedical Optics
• Microscopy
• Tissue Imaging

Background:

• Tissue scattering and refractive index variations limit deep tissue imaging.
• Current clearing techniques improve depth but not necessarily clarity.
• Refractive aberrations, including spherical and Zernike modes, degrade image quality at depth.

Purpose of the Study:

• To investigate the impact of spherical and higher-order aberrations on two-photon microscopy imaging in fixed tissues.
• To compare the effectiveness of correcting only spherical aberration versus correcting the first 22 Zernike aberrations using adaptive optics.

Main Methods:

• Utilized an adaptive optics two-photon microscope.
• Imaged two types of fixed tissue.
• Applied adaptive optics to correct for spherical aberration only and for the first 22 Zernike aberrations.

Main Results:

• Adaptive optics correction significantly improved image quality.
• Compensating for all 22 Zernike aberrations resulted in a 1.6x increase in image intensity.
• Root mean square error was reduced by 3x when correcting for all aberrations.

Conclusions:

• Adaptive optics effectively corrects refractive aberrations in deep tissue imaging.
• Comprehensive aberration correction (22 Zernike modes) offers superior performance over correcting only spherical aberration.
• Adaptive optics enhances imaging clarity and efficiency in biological tissues.