title: Quantitation of fixative-induced morphologic and antigenic variation in mouse and human breast cancers.
abstract: Quantitative Image Analysis (QIA) of digitized whole slide images for morphometric parameters and immunohistochemistry of breast cancer antigens was used to evaluate the technical reproducibility, biological variability, and intratumoral heterogeneity in three transplantable mouse mammary tumor models of human breast cancer. The relative preservation of structure and immunogenicity of the three mouse models and three human breast cancers was also compared when fixed with representatives of four distinct classes of fixatives. The three mouse mammary tumor cell models were an ER+/PR+ model (SSM2), a Her2+ model (NDL), and a triple negative model (MET1). The four breast cancer antigens were ER, PR, Her2, and Ki67. The fixatives included examples of (1) strong cross-linkers, (2) weak cross-linkers, (3) coagulants, and (4) combination fixatives. Each parameter was quantitatively analyzed using modified Aperio Technologies ImageScope algorithms. Careful pre-analytical adjustments to the algorithms were required to provide accurate results. The QIA permitted rigorous statistical analysis of results and grading by rank order. The analyses suggested excellent technical reproducibility and confirmed biological heterogeneity within each tumor. The strong cross-linker fixatives, such as formalin, consistently ranked higher than weak cross-linker, coagulant and combination fixatives in both the morphometric and immunohistochemical parameters.
authors: Center for Comparative Medicine, University of California, Davis, CA 95616, USA.
authors: Robert D
authors: RD
authors: Cardiff
authors: Neil E
authors: NE
authors: Hubbard
authors: Jesse A
authors: JA
authors: Engelberg
authors: Robert J
authors: RJ
authors: Munn
authors: Claramae H
authors: CH
authors: Miller
authors: Judith E
authors: JE
authors: Walls
authors: Jane Q
authors: JQ
authors: Chen
authors: Héctor A
authors: HA
authors: Velásquez-García
authors: Jose J
authors: JJ
authors: Galvez
authors: Katie J
authors: KJ
authors: Bell
authors: Laurel A
authors: LA
authors: Beckett
authors: Yue-Ju
authors: YJ
authors: Li
authors: Alexander D
authors: AD
authors: Borowsky
mesh_terms: D000818:Animals; D000940:Antigenic Variation / Q000187:drug effects*; D014408:Biomarkers, Tumor / Q000032:analysis*; D045744:Cell Line, Tumor; D005260:Female; D005404:Fixatives / Q000494:pharmacology*; D006801:Humans; D007150:Immunohistochemistry; D008099:Liver / Q000187:drug effects; D008325:Mammary Neoplasms, Experimental / Q000473:pathology*; D051379:Mice; D057507:Mice, 129 Strain; D015203:Reproducibility of Results; D013048:Specimen Handling / Q000592:standards*
chemical_list: D014408:Biomarkers, Tumor; D005404:Fixatives
references: Am J Clin Pathol. 2008 Dec;130(6):879-82
references: 19019763
references: Diagn Mol Pathol. 2006 Jun;15(2):115-23
references: 16778593
references: Breast Cancer Res. 2012 Jan 20;14(1):R16
references: 22264274
references: Eur J Clin Chem Clin Biochem. 1997 Aug;35(8):633-5
references: 9298355
references: Biotech Histochem. 2009 Oct;84(5):223-47
references: 19886759
references: J Cell Mol Med. 2008 Oct;12(5B):2027-36
references: 19012729
references: Proteomics Clin Appl. 2009 Aug 1;3(8):874-882
references: 20871745
references: J Natl Compr Canc Netw. 2009 Sep;7 Suppl 6:S1-S21; quiz S22-3
references: 19755043
references: Ann Oncol. 2010 Apr;21(4):717-722
references: 19825882
references: Diagn Mol Pathol. 2010 Jun;19(2):112-22
references: 20502189
references: Cancer Res. 1990 Jun 15;50(12):3545-50
references: 2187598
references: J Mol Diagn. 2012 Sep;14(5):458-66
references: 22749745
references: Lab Invest. 2012 Sep;92(9):1342-57
references: 22801299
references: J Mol Diagn. 2006 May;8(2):157-69
references: 16645201
references: Exp Mol Pathol. 2006 Apr;80(2):183-91
references: 16332367
references: BMC Clin Pathol. 2008 Jan 29;8:1
references: 18230182
references: Evid Rep Technol Assess (Full Rep). 2007 Dec;(160):1-105
references: 18457476
references: Hum Reprod. 1999 Jan;14(1):190-7
references: 10374119
references: Biotech Histochem. 1997 Sep;72(5):273-82
references: 9408588
references: Expert Rev Mol Diagn. 2008 Nov;8(6):707-25
references: 18999923
references: Appl Immunohistochem Mol Morphol. 2008 Mar;16(2):105-7
references: 18227735
references: J Microsc. 2009 Apr;234(1):103-12
references: 19335461
references: J Clin Oncol. 2011 Nov 10;29(32):4273-8
references: 21990413
references: J Histochem Cytochem. 2011 Apr;59(4):356-65
references: 21411807
references: J Histochem Cytochem. 1985 Aug;33(8):845-53
references: 3894502
references: Cytometry A. 2010 Aug;77(8):798-804
references: 20653019
references: Toxicol Pathol. 2011 Oct;39(6):988-1002
references: 21971934
references: Clin Exp Metastasis. 2005;22(1):47-59
references: 16132578
references: J Pathol Inform. 2010 Dec 23;1:29
references: 21221174
references: J Histochem Cytochem. 2008 Nov;56(11):1033-42
references: 18711211
references: Biotech Histochem. 2009 Oct;84(5):217-21
references: 19886758
references: Biotech Histochem. 2008 Dec;83(6):261-77
references: 19031284
references: Lab Invest. 2003 Oct;83(10):1427-35
references: 14563944
references: Arch Pathol Lab Med. 2005 Apr;129(4):502-6
references: 15794674
references: Toxicol Pathol. 2004 Mar-Apr;32(2):181-91
references: 15200156
references: EMBO J. 1999 Apr 15;18(8):2149-64
references: 10205169
references: Nucleic Acids Res. 2007;35(12):e85
references: 17576663
references: Arch Pathol Lab Med. 2011 Jun;135(6):744-52
references: 21631267
references: Am J Pathol. 2002 Dec;161(6):1961-71
references: 12466110
references: J Clin Pathol. 2009 May;62(5):429-34
references: 18818264