Methodology optimization for fluorescence imaging of nervous tissue across in vivo and ex vivo applications

The Gibbs lab has developed a large library of small molecule nerve-specific fluorophores; however, the current resolution of binding is limited by macroscopic fluorescence imaging (>200 nm). Determination of biomolecular interactions will enable medicinal chemistry approaches for novel compounds, off-target binding predictions, and disease-specific applications. Various genetic, biochemical, and computational approaches are ongoing to determine sub-protein level binding, but all will utilize fluorescence imaging as confirmation of fluorophore behavior given the application of fluorophores in Fluorescence Guided Surgery (FGS). Nerve fluorophores are macroscopically imaged in vivo, with higher resolution microscopic techniques relaying heavy tissue processing that introduce artifacts and leached non-covalently bound fluorophore, confounding direct comparisons of fluorophore binding between in vivo and ex vivo imaging. The optimization of methodology for imaging nervous tissue across in vivo and ex vivo (3-D, 2-D) will enable powerful insights into the biomolecular target of nerve-specific fluorophores through high-resolution fluorescence imaging.