Structural, biochemical and biomechanical changes in collagenous tissues upon digestion and cross-linking monitored by combined optical and force microscopy imaging techniques

We demonstrate the capability of linear and non-linear optical microspectroscopy techniques such as fluorescence lifetime imaging (FLIM), Raman spectroscopy (RS), second-harmonic generation (SHG), and two-photon fluorescence (TPF), to monitor structural, biochemical and biomechanical alterations in collagenous tissues upon both collagen digestion and cross-linking using genipin as cross-linker. Thanks to a unique combination of optical and force microscopy techniques in a correlative manner, we obtained detailed information about the biochemical, structural, and biomechanical properties of collagenous tissues upon both digestion and cross-linking treatment.