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Label-free multiphoton imaging to detect hepatic steatosis in chronic liver disease

Hepatic steatosis, the accumulation of lipids within hepatocytes, is defined as intrahepatic fat of at least 5% of liver weight and is an important histological feature. Steatosis may manifest in a variety of liver diseases, and its clinical significance depends on the degree of hepatic steatosis. Excess intrahepatic fat content is a risk factor for disease progression. Increased hepatic steatosis could trigger metabolic dysfunction leading to insulin resistance, dyslipidemia, cardiovascular disease, and progression to non-alcoholic steatohepatitis (NASH), cirrhosis, and hepatocellular carcinoma (HCC). In many chronic liver diseases, hepatic steatosis is associated with increased hepatic fibrosis. Clinical methods of quantifying hepatic steatosis remain semi-quantitative, with potential limitations in precision. Moreover, the evaluation of hepatic steatosis and fibrosis cannot be performed simultaneously. In this work, multiphoton microscopy (MPM) combined two-photon excited fluorescence with second harmonic generation imaging was used to identify the hepatic steatosis and fibrosis in chronic liver disease. The result showed that MPM has the potential to be a pathological diagnostic tool for hepatic steatosis and fibrosis.