Abstract
Conventional histology with light microscopy is essential in the diagnosis of most liver diseases. Recently, a concept of real-time histology with optical biopsy has been advocated. In this study, live mice livers (normal, with fibrosis, steatosis, hepatocellular carcinoma and ischemia-reperfusion injury) were imaged by MPM-FLIM for stain-free real-time histology. The acquired MPM-FLIM images were compared with conventional histological images. MPM-FLIM imaged subsurface cellular and subcellular histopathological hallmarks of live liver in mice models at high resolution. Additional information such as distribution of stellate cell associated autofluorescence and fluorescence lifetime changes was also gathered by MPM-FLIM simultaneously, which cannot be obtained from conventional histology. MPM-FLIM could simultaneously image and quantify the cellular morphology and microenvironment of live livers without conventional biopsy or fluorescent dyes. We anticipate that in the near future MPM-FLIM will be evaluated from bench to bedside, leading to real-time histology and dynamic monitoring of human liver diseases.
Highlights
Liver diseases are frequently encountered in clinical practice with high morbidity and mortality [1]
Conventional histology with light microscopy is essential in the diagnosis of most liver diseases
MPM-FLIM imaged subsurface cellular and subcellular histopathological hallmarks of live liver in mice models at high resolution. Additional information such as distribution of stellate cell associated autofluorescence and fluorescence lifetime changes was gathered by MPM-FLIM simultaneously, which cannot be obtained from conventional histology
Summary
Liver diseases are frequently encountered in clinical practice with high morbidity and mortality [1]. Primary sclerosing cholangitis is a chronic cholestatic liver disease and often develops to liver cirrhosis or even cholangiocarcinoma [3]. Hepatocellular carcinoma is the third leading cause of cancer-related mortality worldwide [4]. Histological examination is essential in the diagnosis of most liver diseases. Conventional histology relies on microscopic examination of a specimen obtained from biopsy or surgery, which is usually time consuming due to the complicated sample preparation procedures including fixation, sectioning and staining. The frozen sections of surgical margin are assessed during hepatocellular carcinoma or cholangiocarcinoma surgery to achieve a R0 resection (complete resection with microscopic examination of margins showing no tumor cells) [5]. Re-resection and/or re-anastomosis must be performed if the surgical margin is tumor positive, leading to the prolonged operation time. There is a desperate need for a rapid, accurate and safe histological diagnosis method for the successful diagnosis and management of liver disease
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