Spectral Multiplexing of Fluorescent Endoscopy for Simultaneous Imaging with Multiple Fluorophores and Multiple Fields of View

Complex clinical procedures and small-animal research procedures can benefit from dual-site imaging provided by multiple endoscopic devices. Here, an endoscopic system is proposed which enables multiple fluorescence microendoscopes to be spectrally multiplexed on a single microscope base, enabling light sources and optical relays to be shared between endoscopes. The presented system is characterized for resolution using USAF-1951 resolution test charts and for modulation transfer function using the slanted edge method. Imaging is demonstrated both directly and with microendoscopes attached. Imaging of phantoms was demonstrated by targeting USAF charts and fiber tissues dyed for FITC and Texas Red fluorescence. Afterwards, simultaneous liver and kidney imaging was demonstrated in mice expressing mitochondrial Dendra2 and injected with Texas Red-dextran. The results indicate that the system achieves high channel isolation and submicron and subcellular resolution, with resolution limited by the endoscopic probe and by physiological movement during endoscopic imaging. Multi-channel microendoscopy provides a potentially low-cost means of simultaneous multiple endoscopic imaging during biological experiments, resulting in reduced animal harm and potentially increasing insight into temporal connections between connected biological systems.

Automated summary

Complex clinical and small-animal research procedures can benefit from dual-site imaging with the proposed endoscopic system, which enables multiple fluorescence microendoscopes to share light sources and optical relays on a single microscope base. The system achieves high channel isolation and submicron and subcellular resolution, as demonstrated by imaging experiments on USAF charts and dyed fiber tissues. Simultaneous liver and kidney imaging was also successfully demonstrated in mice. This multi-channel microendoscopy system provides a potentially low-cost means of simultaneous multiple endoscopic imaging, reducing animal harm and enhancing understanding of temporal connections between biological systems.