A domed window chamber for multi-modality optical imaging

Area of Science:

• Biomedical Engineering
• Microscopy
• Optical Imaging

Background:

• Dorsal skin flap window chambers are essential for in vivo microscopy.
• Current chambers with flat glass windows limit light sheet fluorescence microscopy (LSFM) performance.
• LSFM requires specific sample geometry and refractive index matching for optimal imaging.

Purpose of the Study:

• To engineer a modified window chamber for enhanced multimodal in vivo imaging.
• To adapt dorsal skin flap window chambers for light sheet fluorescence microscopy (LSFM).
• To enable simultaneous optical coherence tomography (OCT), multiphoton microscopy (MPM), and LSFM.

Main Methods:

• Designed a modified window chamber with a domed viewing window made of fluorinated ethylene propylene (FEP).
• FEP material was chosen for its refractive index (n) matching properties similar to water and tissue.
• Validated the chamber design through in vitro imaging of collagen gels and microsphere phantoms.
• Demonstrated in vivo multimodality imaging using OCT, MPM, and LSFM on a custom mouse platform.

Main Results:

• The domed FEP window showed minimal impact on signal strength and image resolution in vitro.
• The modified chamber successfully accommodated LSFM requirements for sample positioning and refractive index.
• Successful in vivo demonstration of simultaneous OCT, MPM, and LSFM imaging was achieved.

Conclusions:

• The modified dorsal skin flap window chamber with a domed FEP window is compatible with LSFM, OCT, and MPM.
• This innovation expands the capabilities of in vivo multimodal imaging in preclinical research.
• The design facilitates advanced optical microscopy techniques for studying biological processes.