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Non-linear multimodal imaging characteristics of early septic liver injury in a mouse model of peritonitis.

Sepsis constitutes a life-threatening organ failure caused by a deregulated host response to infection. Identifying early biomolecular indicators of organ dysfunction may improve clinical decision-making and outcome of patients. Herein, we utilized label-free non-linear multimodal imaging, combining coherent anti-stokes Raman scattering (CARS), two-photon excited auto fluorescence (TPEF) and second-harmonic generation (SHG) to investigate the consequences of early septic liver injury in a murine model of polymicrobial abdominal infection. Liver tissue sections from mice with and without abdominal sepsis were analyzed using multimodal non-linear microscopy, immunofluorescence, immunohistochemistry, and quantitative reverse transcription polymerase chain reaction (qRT-PCR). Twenty-four hours after the induction of sepsis, hepatic mRNA of inflammatory cytokines and acute phase proteins was upregulated, and liver-infiltrating myeloid cells could be visualized alongside hepatocellular cyto-plasmic translocation of high mobility group box 1. According to the statistical analysis based on texture feature extraction fol-lowed by the combination of dimension reduction and linear discriminant analysis, CARS (AUC = 0.93) and TPEF (AUC = 0.83) show an excellent discrimination between liver sections from septic mice and sham-treated mice in contrast to SHG (AUC = 0.49). Spatial analysis revealed no major differences in the distribution of sepsis-associated changes between periportal and pericentral zones. These data suggest early alterations in hepatic lipid distribution and metabolism during liver injury and confirm non-linear multimodal imaging as a promising complementary method for the real-time, label-free study of septic liver damage.