Multimodal optical coherence tomography and two-photon light sheet fluorescence microscopy for embryo imaging

Area of Science:

• Biomedical Engineering
• Developmental Biology
• Microscopy

Background:

• Simultaneous structural and molecular imaging of developing embryos is crucial for understanding developmental pathologies.
• Existing techniques often lack the capability for simultaneous detection of both structural and molecular parameters.

Purpose of the Study:

• To develop a high-resolution, multimodal embryonic imaging system for simultaneous structural and molecular analysis.
• To demonstrate the feasibility and capabilities of the integrated imaging system.

Main Methods:

• Developed a multimodal imaging system by optically co-aligning optical coherence tomography (OCT) and two-photon light sheet fluorescence microscopy (2P-LSFM) beams.
• Utilized galvanometer-mounted mirrors and a shared illumination objective for scanning the beams.
• Achieved specific resolutions: OCT (lateral ~15 µm, axial ~7 µm) and 2P-LSFM (sheet thickness ~10 µm, transverse ~2 µm).

Main Results:

• Successfully demonstrated simultaneous structural and molecular imaging of fluorescent microbeads and fluorescently tagged mouse embryos.
• The integrated system achieved high resolution for both OCT and 2P-LSFM modalities.
• Co-alignment facilitated straightforward image registration.

Conclusions:

• The combined OCT and 2P-LSFM system represents the first successful integration for simultaneous embryonic imaging.
• The co-alignment enables efficient, high-throughput multimodal imaging and simplifies image registration.
• This technology offers significant potential for advancing research in developmental biology and pathology.