Line scanning, fiber bundle fluorescence HiLo endomicroscopy with confocal slit detection.

Area of Science:

• Biomedical Optics
• In Vivo Imaging
• Microscopy

Background:

• Fiber bundle fluorescence endomicroscopy enables in vivo biological tissue imaging.
• Line-scanning confocal laser endomicroscopy offers high frame rates but limited optical sectioning.
• Existing methods struggle to balance imaging speed and axial resolution.

Purpose of the Study:

• To enhance optical sectioning in fiber bundle endomicroscopy while maintaining high acquisition rates.
• To introduce a novel Confocal HiLo endomicroscopy system.
• To evaluate the system's performance against existing techniques.

Main Methods:

• Developed a fiber bundle endomicroscopy system integrating the HiLo technique.
• Synchronized a scanning hybrid-illumination laser line with a CMOS camera's rolling shutter.
• Implemented Confocal HiLo endomicroscopy by combining line-scanning confocal principles with HiLo illumination.
• Tested the system on lens tissue and porcine kidney tissue.

Main Results:

• Achieved significant improvement in axial sectioning compared to standard line-scanning confocal endomicroscopy.
• Demonstrated maintained high image acquisition rates.
• Showcased superior optical sectioning enhancement compared to epifluorescence endomicroscopy, with and without HiLo.
• Validated performance on biological and non-biological tissue samples.

Conclusions:

• Confocal HiLo endomicroscopy effectively enhances optical sectioning in fiber bundle systems.
• The technique successfully balances high-speed imaging with improved axial resolution.
• This advancement offers superior in vivo imaging capabilities for biological tissues.