Second-harmonic patterned polarization-analyzed reflection confocal microscope

Chukwuemeka Okoro, Kimani C. Toussaint

Research output: Contribution to journalArticlepeer-review

Abstract

We introduce the second-harmonic patterned polarization-analyzed reflection confocal (SPPARC) microscope - a multimodal imaging platform that integrates Mueller matrix polarimetry with reflection confocal and second-harmonic generation (SHG) microscopy. SPPARC microscopy provides label-free three-dimensional (3-D), SHG-patterned confocal images that lend themselves to spatially dependent, linear polarimetric analysis for extraction of rich polarization information based on the Mueller calculus. To demonstrate its capabilities, we use SPPARC microscopy to analyze both porcine tendon and ligament samples and find differences in both circular degree-of-polarization and depolarization parameters. Moreover, using the collagen-generated SHG signal as an endogenous counterstain, we show that the technique can be used to provide 3-D polarimetric information of the surrounding extrafibrillar matrix plus cells or EFMC region. The unique characteristics of SPPARC microscopy holds strong potential for it to more accurately and quantitatively describe microstructural changes in collagen-rich samples in three spatial dimensions.

Original languageEnglish (US)
Article number086007
JournalJournal of biomedical optics
Volume22
Issue number8
DOIs
StatePublished - Aug 1 2017

Keywords

  • Mueller matrix
  • confocal microscopy
  • extrafibrillar matrix
  • polarimetry
  • second-harmonic generation

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Biomaterials
  • Atomic and Molecular Physics, and Optics
  • Biomedical Engineering

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