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Second harmonic generation microscopy is a novel technique for differential diagnosis of breast fibroepithelial lesions
  1. Wai Jin Tan1,
  2. Jie Yan1,
  3. Shuoyu Xu1,
  4. Aye Aye Thike2,
  5. Boon Huat Bay3,
  6. Hanry Yu1,4,5,6,
  7. Min-Han Tan1,
  8. Puay Hoon Tan2,3,7
  1. 1Institute of Bioengineering and Nanotechnology, Singapore
  2. 2Department of Pathology, Singapore General Hospital, Singapore
  3. 3Department of Anatomy, Yong Loo Lin School of Medicine, National University of Singapore, Singapore
  4. 4Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore
  5. 5Mechanobiology Institute, National University of Singapore, Singapore
  6. 6Singapore-MIT Alliance for Research and Technology, Singapore
  7. 7Duke-NUS Graduate Medical School, Singapore
  1. Correspondence to Dr Min-Han Tan, Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, #04-01,138669, Singapore; mhtan@ibn.a-star.edu.sg or Dr Puay Hoon Tan, Department of Pathology, Singapore General Hospital, 20 College Road, Academia, Level 7, Diagnostics Tower, 169856, Singapore; tan.puay.hoon{at}sgh.com.sg

Abstract

Breast fibroepithelial lesions, including fibroadenomas and phyllodes tumours, are commonly encountered in clinical practice. As histological differences between these two related entities may be subtle, resulting in a challenging differential diagnosis, pathological techniques to assist the differential diagnosis of these two entities are of high interest. An accurate diagnosis at biopsy is important given corresponding implications for clinical decision-making including surgical extent and monitoring. Second harmonic generation (SHG) microscopy is a recently developed optical imaging technique capable of robust, powerful and unbiased label-free direct detection of collagen fibril structure in tissue without the use of antibodies. We constructed tissue microarrays emulating limited materials on biopsy to investigate quantitative collagen signal in fibroepithelial lesions using SHG microscopy. Archived formalin-fixed paraffin-embedded materials of 47 fibroepithelial lesions (14 fibroadenomas and 33 phyllodes tumours) were evaluated. Higher collagen signal on SHG microscopy was observed in fibroadenomas than phyllodes tumours on SHG imaging (p<0.001, area under the curve 0.859). At an automated threshold (2.5 million positive pixels), the sensitivity and specificity of the SHG microscopy for fibroadenoma classification was 71.4% and 84.4%, respectively. To corroborate these findings, we performed immunohistochemistry on tissue array sections using collagen I and III primary antibodies. Both collagen I and III immunohistochemical expressions were also significantly higher in fibroadenomas than in phyllodes tumours (p<0.001). In conclusion, label-free collagen quantitation on SHG microscopy is a novel imaging approach that can aid the differential diagnosis of fibroepithelial lesions.

  • BREAST PATHOLOGY
  • IMAGE ANALYSIS
  • IMMUNOHISTOCHEMISTRY

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