Mitigating fluorescence spectral overlap in wide-field endoscopic imaging.

Area of Science:

• Biomedical optics
• Molecular imaging
• Fluorescence spectroscopy

Background:

• Multispectral fluorescence imaging is crucial for molecular detection but limited by overlapping fluorophore emission spectra.
• Fluorophore emission cross-talk hinders accurate signal detection and analysis in molecular imaging.

Purpose of the Study:

• To evaluate methods for removing fluorophore emission cross-talk in wide-field multispectral fluorescence molecular imaging.
• To assess the viability of these methods using a scanning fiber endoscope (SFE).

Main Methods:

• Fabrication of a phantom with controlled fluorophore emission cross-talk.
• Testing three cross-talk removal techniques: image stitching, concurrent imaging with cross-talk ratio subtraction, and frame-sequential imaging.
• Utilizing a 1.2-mm ultrathin scanning fiber endoscope (SFE) for in vivo testing.

Main Results:

• Successful avoidance or significant reduction of fluorophore emission cross-talk was achieved.
• Concurrent imaging with cross-talk subtraction demonstrated viability for in vivo applications.
• Demonstrated enhancement of exogenous fluorescence target-to-background ratio by reducing tissue autofluorescence.

Conclusions:

• Fluorophore emission cross-talk can be effectively managed in multispectral fluorescence molecular imaging.
• Concurrent imaging with cross-talk subtraction using SFE is a promising approach for early cancer detection and localization.
• Techniques for reducing autofluorescence background can improve imaging sensitivity.