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Related Experiment Video

Updated: Feb 24, 2026

Author Spotlight: Non-Invasive Imaging of Complex Bio-Structures Using Polarization-Sensitive Two-Photon Microscopy

Author Spotlight: Non-Invasive Imaging of Complex Bio-Structures Using Polarization-Sensitive Two-Photon Microscopy

Published on: September 8, 2023

Second-harmonic patterned polarization-analyzed reflection confocal microscope.

Chukwuemeka Okoro¹, Kimani C Toussaint^2,3

¹University of Illinois at Urbana-Champaign, PROBE Lab, Department of Electrical and Computer Enginee, United States.

Journal of Biomedical Optics

|August 25, 2017

Summary

This summary is machine-generated.

We developed a new multimodal imaging microscope called SPPARC microscopy. This technique analyzes collagen-rich tissues, revealing microstructural differences in three dimensions using label-free polarization and SHG imaging.

Keywords:

Mueller matrix confocal microscopy extrafibrillar matrix polarimetry second-harmonic generation

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Area of Science:

Biomedical Optics
Materials Science
Microscopy

Background:

Collagen-rich tissues exhibit complex microstructures crucial for their function.
Label-free imaging techniques are needed for quantitative microstructural analysis.
Mueller matrix polarimetry and second-harmonic generation (SHG) microscopy offer complementary information.

Purpose of the Study:

To introduce and validate the second-harmonic patterned polarization-analyzed reflection confocal (SPPARC) microscope.
To demonstrate SPPARC microscopy's capability for label-free 3-D microstructural analysis of collagenous tissues.
To extract detailed polarization information using Mueller calculus.

Main Methods:

Integration of Mueller matrix polarimetry, reflection confocal microscopy, and SHG microscopy into a single platform (SPPARC).
Acquisition of SHG-patterned confocal images for spatially dependent polarimetric analysis.
Analysis of porcine tendon and ligament samples.

Main Results:

SPPARC microscopy provides label-free 3-D SHG-patterned confocal images.
Spatially dependent polarimetric analysis revealed differences in circular degree-of-polarization and depolarization parameters between tendon and ligament.
SHG signal from collagen enabled 3-D polarimetric mapping of the extrafibrillar matrix plus cells (EFMC) region.

Conclusions:

SPPARC microscopy is a powerful multimodal imaging platform for label-free 3-D analysis of collagen-rich tissues.
The technique quantitatively describes microstructural changes by combining polarimetric and SHG information.
SPPARC microscopy holds significant potential for studying diseases affecting collagenous tissues.