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A Standardized Method for the Analysis of Liver Sinusoidal Endothelial Cells and Their Fenestrations by Scanning Electron Microscopy
Published on: April 30, 2015
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Imaging fenestrations in liver sinusoidal endothelial cells by optical localization microscopy
Viola Mönkemöller1, Mark Schüttpelz, Peter McCourt
1Biomolecular Photonics, Department of Physics, University of Bielefeld, 33615 Bielefeld, Germany. thomas.huser@physik.uni-bielefeld.de.
Physical Chemistry Chemical Physics : PCCP
|May 17, 2014
Summary
Direct stochastic optical reconstruction microscopy (dSTORM) successfully visualized liver sinusoidal endothelial cell (LSEC) fenestrations, overcoming optical limits. This breakthrough enables detailed study of these crucial cells without electron microscopy.
Area of Science:
- Cell Biology
- Microscopy Techniques
- Hepatology
Background:
- Liver sinusoidal endothelial cells (LSECs) are vital for liver function, regulating transport between blood and liver.
- LSEC dysfunction is linked to metabolic and aging disorders.
- Investigating LSECs is challenging due to the small size of their fenestrations (50-200 nm).
Purpose of the Study:
- To resolve the ultrastructure of LSEC fenestrations using super-resolution microscopy.
- To compare the effectiveness of dSTORM with 3D-SIM for imaging LSEC fenestrations.
- To establish a method for studying LSECs in biomedical laboratories.
Main Methods:
- Direct stochastic optical reconstruction microscopy (dSTORM) was employed to achieve high spatial resolution (∼20 nm).
- Cellular plasma membranes were labeled with CellMask Deep Red at high fluorophore density.
- Imaging was performed using a reducing buffer system, and results were compared to 3D structured illumination microscopy (3D-SIM).
Main Results:
- dSTORM resolved LSEC fenestrations with unprecedented spatial resolution of approximately 20 nm.
- The study demonstrated the superior structural detail provided by dSTORM compared to 3D-SIM.
- The method allows for detailed imaging of LSEC fenestrations, crucial for understanding cell function.
Conclusions:
- dSTORM provides a powerful tool for visualizing LSEC fenestrations below the optical diffraction limit.
- This technique offers a viable alternative to electron microscopy for studying LSECs.
- The findings facilitate broader investigation of LSEC function in various physiological and pathological conditions.
