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In vivo imaging of hepatic excretory function in the rat by fluorescence microscopy
Peter Recknagel1, Ralf A Claus, Ute Neugebauer
1Integrated Research and Treatment Center-Center for Sepsis Control and Care-CSCC, Jena University Hospital, Erlanger Allee 101, 07747 Jena, Germany.
Abstract:
Applying intravital fluorescence microscopy, we assessed sinusoidal delivery and biliary clearance of two different polymethine dyes. DY635, a benzopyrylium-based hemocyanine dye with shorter excitation wavelength than indocyanine green (ICG), was validated for assessment of hepatic excretory function. Decrease of DY635 and ICG reflecting transcellular transport was 83 ± 4% (DY635) and 14 ± 2% (ICG; p < 0.05) over 35 minutes, respectively. In cholestasis, hepatobiliary excretion of DY635 was markedly impaired (control 3176 ± 148 pmol vs. cholestatic 1929 ± 179 pmol; p < 0.05). DY635 even enabled an analysis at high resolution suggesting 1.) hepatocyte uncoupling and 2.) failure of primarily the canalicular pole, allowing in vivo insights into molecular mechanisms of this critical facet of hepatobiliary function.
Insights
This study validates DY635, a novel hemocyanine dye, for assessing liver function. DY635 offers superior insights into hepatobiliary excretion compared to indocyanine green, especially in cholestasis models.
Area of Science:
- Hepatology and Biomedical Imaging
- Molecular Mechanisms of Liver Excretion
Background:
- Intravital fluorescence microscopy was employed to evaluate the sinusoidal delivery and biliary clearance of polymethine dyes.
- DY635, a benzopyrylium-based hemocyanine dye, was assessed for its utility in evaluating hepatic excretory function, offering a shorter excitation wavelength than indocyanine green (ICG).
Discussion:
- DY635 demonstrated significantly higher transcellular transport compared to ICG (83% vs. 14% over 35 minutes).
- In a cholestasis model, DY635 excretion was markedly impaired, indicating its sensitivity to altered hepatobiliary function.
- The dye facilitated high-resolution analysis, revealing potential hepatocyte uncoupling and canalicular pole dysfunction.
Key Insights:
- DY635 serves as a sensitive and effective tool for assessing hepatic excretory function in vivo.
- The study provides novel insights into the molecular mechanisms underlying impaired biliary clearance in cholestasis.
- High-resolution imaging with DY635 allows for detailed analysis of hepatocyte and canalicular transport.
Outlook:
- DY635 has the potential to advance the diagnosis and understanding of liver diseases characterized by impaired biliary excretion.
- Further research can explore DY635's application in other liver function assessments and drug development.
- This imaging approach may offer a non-invasive method to study liver pathophysiology at a molecular level.
