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Simultaneous Label-Free Autofluorescence Multi-Harmonic Microscopy
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Label-free imaging of bone multiscale porosity and interfaces using third-harmonic generation microscopy.
Rachel Genthial1,2, Emmanuel Beaurepaire3, Marie-Claire Schanne-Klein3
1Univ. Grenoble Alpes, LIPHY, F-38000, Grenoble, France.
Scientific Reports
|June 15, 2017
Summary
New label-free microscopy visualizes bone
Area of Science:
- Biomedical Engineering
- Materials Science
- Cell Biology
Background:
- Bone's hierarchical structure, from nano- to millimeters, dictates its function.
- Existing imaging techniques struggle to capture the integration of bone's complex heterogeneities.
- Understanding these structures is crucial for bone physiology and pathology.
Purpose of the Study:
- To introduce a novel label-free microscopy technique for bone imaging.
- To visualize the intricate lacuno-canalicular network (LCN) and remodeling interfaces in 3D.
- To correlate LCN structure with extracellular matrix organization.
Main Methods:
- Utilized label-free third-harmonic generation (THG) microscopy.
- Achieved submicron precision over millimetric fields of view.
- Simultaneously recorded THG and second-harmonic generation (SHG) signals.
Main Results:
- Directly visualized the 3D porous lacuno-canalicular network (LCN) housing osteocytes.
- Identified interfaces corresponding to bone remodeling stages.
- Enabled analysis of LCN structure relative to collagen organization and larger structures.
Conclusions:
- THG microscopy offers unprecedented 3D visualization of bone's microarchitecture.
- This technique overcomes limitations of current imaging methods for bone research.
- Provides a powerful tool for studying bone structure-function relationships.
