Human hepatocellular carcinoma diagnosis by multiphoton autofluorescence microscopy

Abstract

Conventionally, the diagnosis of hepatocellular carcinoma (HCC) is performed by qualitative examination of histopathological specimens, which takes times for sample preparation in fixation, section and stain. Our objective is to demonstrate an effective and efficient approach to apply multiphoton microscopy imaging the HCC specimens, with the advantages of being optical section, label-free, subcellular resolution, minimal invasiveness, and the acquisition of quantitative information at the same time. The imaging modality of multiphoton autofluorescence (MAF) was used for the qualitative imaging and quantitative analysis of HCC of different grades under ex-vivo, label-free conditions. We found that while MAF is effective in identifying cellular architecture in the liver specimens, and obtained quantitative parameters in characterizing the disease. Our results demonstrates the capability of using tissue quantitative parameters of multiphoton autofluorescence (MAF), the nuclear number density (NND), and nuclear-cytoplasmic ratio (NCR) for tumor discrimination and that this technology has the potential in clinical diagnosis of HCC and the in-vivo investigation of liver tumor development in animal models.