Real-time histology in liver disease using multiphoton microscopy with fluorescence lifetime imaging

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 ischemiareperfusion 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.

Fiber‐based fluorescence lifetime imaging of recellularization processes on vascular tissue constructs

“…In addition to their spectral properties, NADH and FAD have fluorescent decay lifetimes that vary according to their protein‐binding state. Consequently, NADH and FAD label‐free FLIm microscopy has been extensively utilized for investigations of metabolic signatures of cancerous cells , to identify histological features of livers , or to probe skin constituents . Conventional approaches to FLIm are based on imaging through microscope objectives, which can achieve high spatial resolution of cells in planar scaffolds or excised tissue, but are not suitable to more challenging geometries such as in situ intravascular imaging.…”

Two-photon dual imaging platform for in vivo monitoring cellular oxidative stress in liver injury

“…The cellular details of the liver can be imaged deep to 250 μm below the fibrous capsule of Glisson using multiphoton microscopy. According to our previous study, imaging depth of 50 to 100 μm was the clearest for observing cellular and subcellular morphology in the liver2445. Hepatocytes around the central vein (Zone 3) are more sensitive to APAP-induced injury than those around the portal vein (Zone 1)46.…”

“…Body temperature was controlled by placing mice on a heating pad set to 37 °C. Intravital imaging of the mouse liver was performed as previously described2445. Briefly, a midline laparotomy is performed and the liver is exposed for imaging.…”

Applications of fluorescence lifetime imaging in clinical medicine

“…FLIM can image and quantify cell morphology and probe the liver microenvironment without conventional biopsy or°uorescent dye. Wang et al 47 believe that FLIM will be used from the laboratory to the clinic for real-time histology and dynamic monitoring of human liver disease in the near future. In 1999, Mizeret et al 48 developed the video frequency domain FLIM in vivo and applied it in 46 (Permission has been granted by \Biomedical Optics Express".)…”