Single source CARS-based multimodal microscopy system for biological tissue imaging [Invited].

Area of Science:

• Biomedical Optics
• Microscopy Techniques
• Advanced Imaging

Background:

• Multimodal microscopy offers complementary information for biological sample analysis.
• Coherent Anti-Stokes Raman Scattering (CARS) microscopy provides label-free chemical contrast.
• Integrating multiple imaging modalities enhances diagnostic and research capabilities.

Purpose of the Study:

• To develop and demonstrate a novel CARS-based multimodal microscopy system.
• To integrate CARS, multiphoton microscopy (MPM), and reflectance confocal microscopy (RCM).
• To achieve high-resolution, co-registered imaging of biological tissues.

Main Methods:

• Utilized a single Ti:sapphire femtosecond laser source.
• Implemented spectral selection for vibrational excitation (approx. 2850 cm⁻¹) targeting lipid cells.
• Employed adjustable spectral filters for tunable excitation.
• Achieved imaging speeds of 1 frame-per-second at 1024 × 1024 pixel resolution.

Main Results:

• Successfully generated clear 2D and 3D images of ex-vivo biological samples.
• Demonstrated co-registered imaging across CARS, MPM, and RCM modalities.
• Validated the system's performance using porcine fat, murine skin, and murine liver tissues.

Conclusions:

• The developed CARS-based multimodal microscopy system is effective for high-resolution biological imaging.
• The system's ability to combine multiple modalities offers comprehensive insights into tissue structures and composition.
• This technology holds potential for advanced research in pathology and biomedical diagnostics.