Wavefront correction with image-based interferometric focus sensing in two-photon microscopy

Area of Science:

• Biomedical Optics
• Microscopy
• Adaptive Optics

Background:

• Adaptive optics (AO) enhances image quality by correcting wavefront distortions.
• Interferometric focus sensing (IFS) is an AO technique effective for deep tissue imaging but typically uses a single sample point.
• Complex aberrations limit imaging depth and clarity in biological samples.

Purpose of the Study:

• To introduce and validate an image-based interferometric focus sensing (IBIFS) method.
• To enable wavefront correction over an extended field of view in microscopy.
• To improve the stability and effectiveness of adaptive optics for deep tissue imaging.

Main Methods:

• Developed an image-based interferometric focus sensing (IBIFS) technique.
• Implemented IBIFS within a sample conjugate adaptive optics configuration.
• Utilized image quality metrics for progressive wavefront estimation and correction.
• Experimentally validated using two-photon microscopy on fluorescent beads and mouse brain slices.

Main Results:

• Demonstrated effective wavefront correction across a large field of view.
• Achieved more stable optimization results compared to region-of-interest based methods.
• Successfully applied IBIFS to complex biological samples like mouse brain slices.

Conclusions:

• IBIFS offers a robust and effective solution for aberration correction in deep tissue imaging.
• The proposed method expands the field of view for adaptive optics microscopy.
• IBIFS provides enhanced stability and performance for advanced imaging applications.