Characterization of pathological thyroid tissue using polarization-sensitive second harmonic generation microscopy


Polarization-sensitive second harmonic generation (SHG) microscopy is an established imaging technique able to provide information related to specific molecular structures including collagen. In this investigation, polarization-sensitive SHG microscopy was used to investigate changes in the collagen ultrastructure between histopathology slides of normal and diseased human thyroid tissues including follicular nodular disease, Grave’s disease, follicular variant of papillary thyroid carcinoma, classical papillary thyroid carcinoma, insular or poorly differentiated carcinoma, and anaplastic or undifferentiated carcinoma ex vivo. The second-order nonlinear optical susceptibility tensor component ratios, χ(2)zzz′/χ(2)zxx′ and χ(2)xyz′/χ(2)zxx′, were obtained, where χ(2)zzz′/χ(2)zxx′ is a structural parameter and χ(2)xyz′/χ(2)zxx′ is a measure of the chirality of the collagen fibers. Furthermore, the degree of linear polarization (DOLP) of the SHG signal was measured. A statistically significant increase in χ(2)zzz′/χ(2)zxx′ values for all the diseased tissues except insular carcinoma and a statistically significant decrease in DOLP for all the diseased tissues were observed compared to normal thyroid. This finding indicates a higher ultrastructural disorder in diseased collagen and provides an innovative approach to discriminate between normal and diseased thyroid tissues that is complementary to standard histopathology.

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Fig. 1: A schematic of the nonlinear optical microscope.
Fig. 2: PIPO SHG microscopy of normal and different pathological thyroid tissues.


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This research was supported by the Natural Sciences and Engineering Research Council of Canada (NSERC RGPIN-2017-06923 and DGDND-2017-00099) and the Canadian Institutes of Health Research (CIHR) through a Collaborative Health Research Project (CHRP) grant (CPG-134752 and CHRPJ 462842-14). The authors thank the staff of the Advanced Optical Microscopy Facility at University Health Network for assistance with whole-slide scanning.

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Correspondence to Danielle Tokarz or Brian C. Wilson or Virginijus Barzda.

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Tokarz, D., Cisek, R., Joseph, A. et al. Characterization of pathological thyroid tissue using polarization-sensitive second harmonic generation microscopy. Lab Invest (2020).

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