Multiphoton microscopy and fluorescence lifetime imaging provide a novel method in studying drug distribution and metabolism in the rat liver in vivo

Thorling, Camilla A.; Dancik, Yuri; Hupple, Clinton; Medley, Gregory; Liu, Xin; Zvyagin, Andrei V.; Robertson, Thomas A.; Burczynski, Frank J.; Roberts, Michael S.

doi:10.1117/1.3614473

Cited by 21 publications

(27 citation statements)

References 42 publications

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“…[34][35][36][37][38][39][40][41][42] As outlined below, we have adapted and extended the approaches used in these studies into quantitative assays of hepatocyte transport.…”

Section: Resultsmentioning

confidence: 99%

Quantitative intravital microscopy of hepatic transport

et al. 2012

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“…[34][35][36][37][38][39][40][41][42] As outlined below, we have adapted and extended the approaches used in these studies into quantitative assays of hepatocyte transport.…”

Section: Resultsmentioning

confidence: 99%

Quantitative intravital microscopy of hepatic transport

et al. 2012

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“…To further confirm these results, we tested our previously published model of measuring average fluorescence lifetime to determine whether metabolism of fluorescein to FG was increased (Thorling et al, 2011a). Fluorescein and FG have two distinct lifetimes, but our analysis yielded one single average lifetime (t m ), which is a combination of the lifetime of fluorescein and FG.…”

Section: Discussionmentioning

confidence: 99%

“…High-Performance Liquid Chromatography. Concentrations of fluorescein and FG in each collected bile sample were determined by high-performance liquid chromatography analysis (Shimadzu, Kyoto, Japan) as previously described elsewhere (Thorling et al, 2011a). In brief, the mobile phase consisted of 50 mM NaH 2 PO 4 and methanol (2:3, v/v).…”

Section: Methodsmentioning

confidence: 99%

“…Fluorescein and FG have overlapping excitation and emission spectra but different fluorescence lifetimes, which makes fluorescence lifetime imaging to detect these lifetime changes possible. Our group has shown that fluorescence lifetime imaging microscopy (FLIM) can study the metabolism of fluorescein to FG (Thorling et al, 2011a). In this study, we used clinically approved fluorescein to examine liver subcellular function changes induced by liver disease.…”

Section: Introductionmentioning

confidence: 99%

“…We used a physiologically-based pharmacokinetic model to simultaneously fit biliary excretion and imaging data after injection of fluorescein. FLIM was used to validate the model, by studying the metabolism of fluorescein to FG as previously described elsewhere (Thorling et al, 2011a). This is, to our knowledge, the first study to quantitate subcellular functional changes in steatotic livers after I/R injury using minimally invasive multiphoton microscopy (MPM).…”

Section: Introductionmentioning

confidence: 99%

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Assessing Steatotic Liver Function after Ischemia-Reperfusion Injury by In Vivo Multiphoton Imaging of Fluorescein Disposition

et al. 2014

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Ischemia-reperfusion injury, a common complication during liver surgery where steatotic livers are more prone to the injury, may become more prevalent in the growing obese population. This study characterizes liver morphology toward understanding changes in subcellular function in steatotic livers exposed to ischemiareperfusion injury through quantitative description of fluorescein distribution obtained by minimally invasive in vivo multiphoton microscopy using a physiologic pharmacokinetic model. Rats were fed a high-fat diet for 7 days to induce liver steatosis. Partial ischemia was induced after reperfusion for 4 hours, when fluorescein (10 mg/kg) was injected intravenously. Liver images, bile, and blood were collected up to 180 minutes after injection. Ischemia-reperfusion injury was associated with an increase in alanine transaminase levels and apoptosis. In addition, steatosis featured lipid droplets and an increase in fluorescein-associated fluorescence observed in hepatocytes via multiphoton imaging. Analysis of the hepatic concentrationtime profiles has suggested that the steatosis-induced increase in fluorescein-associated fluorescence mainly arises by inducing hepatic fluorescein metabolism. The combination of ischemiareperfusion with steatosis exacerbates these effects further. This was confirmed by fluorescence lifetime imaging microscopy showing a decreased average fluorescence lifetime of the liver, which is indicative of an increased production of the metabolite. Our results show the potential of noninvasive dye imaging for improving our understanding of liver disease induced by subcellular changes in vivo, providing further quantitative measures of metabolic and biliary liver function, and hence extending the qualitative liver function tests now available.

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Visualizing liver anatomy, physiology and pharmacology using multiphoton microscopy

Wang

Liang

Gravot

et al. 2016

Journal of Biophotonics

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Multiphoton microscopy (MPM) has become increasingly popular and widely used in both basic and clinical liver studies over the past few years. This technology provides insights into deep live tissues with less photobleaching and phototoxicity, which helps us to better understand the cellular morphology, microenvironment, immune responses and spatiotemporal dynamics of drugs and therapeutic cells in the healthy and diseased liver. This review summarizes the principles, opportunities, applications and limitations of MPM in hepatology. A key emphasis is on the use of fluorescence lifetime imaging (FLIM) to add additional quantification and specificity to the detection of endogenous fluorescent species in the liver as well as exogenous molecules and nanoparticles that are applied to the liver in vivo. We anticipate that in the near future MPM‐FLIM will advance our understanding of the cellular and molecular mechanisms of liver diseases, and will be evaluated from bench to bedside, leading to real‐time histology of human liver diseases.

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Multiphoton microscopy and fluorescence lifetime imaging provide a novel method in studying drug distribution and metabolism in the rat liver in vivo

Cited by 21 publications

References 42 publications

Quantitative intravital microscopy of hepatic transport

Quantitative intravital microscopy of hepatic transport

Assessing Steatotic Liver Function after Ischemia-Reperfusion Injury by In Vivo Multiphoton Imaging of Fluorescein Disposition

Visualizing liver anatomy, physiology and pharmacology using multiphoton microscopy

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