A large-field polarisation-resolved laser scanning microscope: applications to CARS imaging

Area of Science:

• Optical Microscopy
• Biomedical Imaging
• Spectroscopy

Background:

• Laser-scanning imaging techniques rely on excitation light polarization to determine molecular orientation.
• Precise polarization control at the sample is crucial for these techniques.
• Polarization distortions from optical elements limit the field-of-view in polarization-resolved (PR) microscopy.

Purpose of the Study:

• To implement large-field-of-view PR microscopy with suppressed polarization distortions.
• To demonstrate the effectiveness of a carefully designed laser-beam optical path.
• To enable high-quality PR CARS imaging of biological samples, such as mouse spinal cord.

Main Methods:

• Development and implementation of a novel laser-beam optical path design for PR microscopy.
• Quantitative analysis of polarization distortion suppression.
• Application of the technique to Coherent Anti-Stokes Raman Scattering (CARS) imaging of mouse spinal cord.

Main Results:

• Successfully demonstrated large-field-of-view PR microscopy with significantly reduced polarization distortions.
• Quantified the minimized effects of distortions on the final images.
• Achieved high-resolution PR CARS images of mouse spinal cord, revealing detailed structures.

Conclusions:

• The designed optical path effectively suppresses polarization distortions in PR laser-scanning microscopy.
• This approach enables large-field-of-view imaging without compromising image quality.
• The methodology is applicable to various PR laser-scanning techniques beyond CARS imaging.