title: Comparison of Picrosirius Red Staining With Second Harmonic Generation Imaging for the Quantification of Clinically Relevant Collagen Fiber Features in Histopathology Samples.
abstract: Stromal collagen alignment has been shown to have clinical significance in a variety of cancers and in other diseases accompanied by fibrosis. While much of the biological and clinical importance of collagen changes has been demonstrated using second harmonic generation (SHG) imaging in experimental settings, implementation into routine clinical pathology practice is currently prohibitive. To translate the assessment of collagen organization into routine pathology workflow, a surrogate visualization method needs to be examined. The objective of the present study was to quantitatively compare collagen metrics generated from SHG microscopy and commonly available picrosirius red stain with standard polarization microscopy (PSR-POL). Each technique was quantitatively compared with established image segmentation and fiber tracking algorithms using human pancreatic cancer as a model, which is characterized by a pronounced stroma with reorganized collagen fibers. Importantly, PSR-POL produced similar quantitative trends for most collagen metrics in benign and cancerous tissues as measured by SHG. We found it notable that PSR-POL detects higher fiber counts, alignment, length, straightness, and width compared with SHG imaging but still correlates well with SHG results. PSR-POL may provide sufficient and additional information in a conventional clinical pathology laboratory for certain types of collagen quantification.
authors.affiliation: Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, Wisconsin (CRD, AK, WJK, KWE)|Laboratory for Optical and Computational Instrumentation, University of Wisconsin-Madison, Madison, Wisconsin (LOCI) (CRD, KM, GM, AK, YL, WJK, KWE),|Morgridge Institute for Research, Madison, Wisconsin (CRD, KWE)
authors.forename: Cole R
authors.initials: CR
authors.lastname: Drifka
authors.affiliation: Department of Surgical Pathology, University of Wisconsin-Madison, Madison, Wisconsin (AGL)|University of Wisconsin Comprehensive Carbone Cancer Center, Madison, Wisconsin (AGL, WAR, SMW, WJK, KWE)
authors.forename: Agnes G
authors.initials: AG
authors.lastname: Loeffler
authors.affiliation: Laboratory for Optical and Computational Instrumentation, University of Wisconsin-Madison, Madison, Wisconsin (LOCI) (CRD, KM, GM, AK, YL, WJK, KWE),
authors.forename: Kara
authors.initials: K
authors.lastname: Mathewson
authors.affiliation: Laboratory for Optical and Computational Instrumentation, University of Wisconsin-Madison, Madison, Wisconsin (LOCI) (CRD, KM, GM, AK, YL, WJK, KWE),
authors.forename: Guneet
authors.initials: G
authors.lastname: Mehta
authors.affiliation: Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, Wisconsin (CRD, AK, WJK, KWE)|Laboratory for Optical and Computational Instrumentation, University of Wisconsin-Madison, Madison, Wisconsin (LOCI) (CRD, KM, GM, AK, YL, WJK, KWE),
authors.forename: Adib
authors.initials: A
authors.lastname: Keikhosravi
authors.affiliation: Laboratory for Optical and Computational Instrumentation, University of Wisconsin-Madison, Madison, Wisconsin (LOCI) (CRD, KM, GM, AK, YL, WJK, KWE),
authors.forename: Yuming
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Urology, University of Wisconsin-Madison, Madison, Wisconsin (SL, WAR)
authors.forename: Stephanie
authors.initials: S
authors.lastname: Lemancik
authors.affiliation: Department of Urology, University of Wisconsin-Madison, Madison, Wisconsin (SL, WAR)|George M. O'Brien Research Center of Excellence, Madison, Wisconsin (WAR)|University of Wisconsin Comprehensive Carbone Cancer Center, Madison, Wisconsin (AGL, WAR, SMW, WJK, KWE)
authors.forename: William A
authors.initials: WA
authors.lastname: Ricke
authors.affiliation: Department of Surgery, University of Wisconsin-Madison, Madison, Wisconsin (SMW, WJK)|University of Wisconsin Comprehensive Carbone Cancer Center, Madison, Wisconsin (AGL, WAR, SMW, WJK, KWE)
authors.forename: Sharon M
authors.initials: SM
authors.lastname: Weber
authors.affiliation: Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, Wisconsin (CRD, AK, WJK, KWE)|Laboratory for Optical and Computational Instrumentation, University of Wisconsin-Madison, Madison, Wisconsin (LOCI) (CRD, KM, GM, AK, YL, WJK, KWE),|Department of Surgery, University of Wisconsin-Madison, Madison, Wisconsin (SMW, WJK)|University of Wisconsin Comprehensive Carbone Cancer Center, Madison, Wisconsin (AGL, WAR, SMW, WJK, KWE)
authors.forename: W John
authors.initials: WJ
authors.lastname: Kao
authors.affiliation: Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, Wisconsin (CRD, AK, WJK, KWE)|Laboratory for Optical and Computational Instrumentation, University of Wisconsin-Madison, Madison, Wisconsin (LOCI) (CRD, KM, GM, AK, YL, WJK, KWE),|University of Wisconsin Comprehensive Carbone Cancer Center, Madison, Wisconsin (AGL, WAR, SMW, WJK, KWE)|Morgridge Institute for Research, Madison, Wisconsin (CRD, KWE)
authors.forename: Kevin W
authors.initials: KW
authors.lastname: Eliceiri
mesh_terms: D001391:Azo Compounds / Q000737:chemistry*; D004396:Coloring Agents / Q000737:chemistry*; D024022:Fibrillar Collagens / Q000032:analysis*; D006801:Humans; D008853:Microscopy / Q000379:methods; D010190:Pancreatic Neoplasms / Q000737:chemistry*; D046888:Tissue Array Analysis
chemical_list: D001391:Azo Compounds; D004396:Coloring Agents; D024022:Fibrillar Collagens; C009798:C.I. direct red 80
keywords: fibrillar collagen; histopathology; picrosirius red; polarized microscopy; second harmonic generation