Multimodal label‐free imaging of murine hepatocellular carcinoma with a subcellular resolution

We demonstrate label‐free imaging of genetically induced hepatocellular carcinoma (HCC) in a murine model provided by two‐ and three‐photon fluorescence microscopy of endogenous fluorophores excited at the central wavelengths of 790, 980 and 1250 nm and reinforced by second and third harmonic generation microscopy. We show, that autofluorescence imaging presents abundant information about cell arrangement and lipid accumulation in hepatocytes and hepatic stellate cells (HSCs), harmonics generation microscopy provides a versatile tool for fibrogenesis and steatosis study. Multimodal images may be performed by a single ultrafast laser source at 1250 nm falling in tissue transparency window. Various grades of HCC are examined revealing fibrosis, steatosis, liver cell dysplasia, activation of HSCs and hepatocyte necrosis, that shows a great ability of multimodal label‐free microscopy to intravital visualization of liver pathology development. Hepatocellular carcinoma (HCC) in a murine model is stain‐free visualized by two‐, three‐photon autofluorescence and harmonic generation microscopy. Various grades of HCC are examined by multimodal imaging revealing liver cell dysplasia, hepatocyte necrosis, steatosis and fibrosis development by a single ultrafast laser source at the wavelength of 1250 nm.