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A robust collagen scoring method for human liver fibrosis by second harmonic microscopy.
Thomas Guilbert1, Christophe Odin, Yann Le Grand
1Institut of Physics of Rennes IPR/UMR CNRS 6251, University of Rennes I, Campus de Beaulieu, Bat 11A, 35042 Rennes Cedex, France.
Optics Express
|December 18, 2010
Summary
A new collagen scoring method using Second Harmonic Generation (SHG) microscopy is robust for liver fibrosis assessment. This non-invasive technique accurately quantifies collagen without staining, aiding diagnosis and treatment evaluation.
Area of Science:
- Biomedical Imaging
- Histopathology
- Optical Microscopy
Background:
- Second Harmonic Generation (SHG) microscopy enables label-free imaging of Type I collagen.
- Previous work established a correlation between SHG-based collagen scoring and liver fibrosis staging (Metavir).
Purpose of the Study:
- To conduct a detailed investigation of the SHG-based collagen scoring method using two distinct objective lenses.
- To assess the robustness of the scoring method against variations in imaging parameters.
Main Methods:
- Utilized measurements of objective point spread functions and photomultiplier gain.
- Employed a simplified model of SHG intensity.
- Applied the scoring method to histological human liver biopsies.
- Evaluated robustness against numerical aperture (NA), reference sample, laser power, and spatial sampling.
Main Results:
- The SHG collagen scoring method demonstrated robustness across different low NA objectives.
- The method proved insensitive to variations in reference sample, laser power, and spatial sampling rate.
- Consistent correlation with fibrosis levels was maintained under tested conditions.
Conclusions:
- The developed collagen scoring method is simple and robust for quantifying collagen in human liver biopsies.
- This technique holds potential for broader applications in assessing collagen content in various organs.
- It can provide valuable diagnostic information and aid in evaluating therapeutic efficacy.
