Quantitation of fixative-induced morphologic and antigenic variation in mouse and human breast cancers

Robert Cardiff, Neil Hubbard, Jesse A. Engelberg, Robert J. Munn, Claramae H. Miller, Judith E. Walls, Jane Q. Chen, Héctor A. Velásquez-García, Jose J. Galvez, Katie J. Bell, Laurel A Beckett, Yue Ju Li, Alexander D Borowsky

Research output: Contribution to journalArticle

29 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)480-497
Number of pages18
JournalLaboratory Investigation
Volume93
Issue number4
DOIs
StatePublished - Apr 2013

Keywords

  • estrogen receptor
  • Her2
  • IHC4 biomarker panel
  • immunohistochemistry
  • MIB1 (Ki67)
  • progesterone receptor
  • quantitative image analysis

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Cell Biology
  • Molecular Biology

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    Cardiff, R., Hubbard, N., Engelberg, J. A., Munn, R. J., Miller, C. H., Walls, J. E., Chen, J. Q., Velásquez-García, H. A., Galvez, J. J., Bell, K. J., Beckett, L. A., Li, Y. J., & Borowsky, A. D. (2013). Quantitation of fixative-induced morphologic and antigenic variation in mouse and human breast cancers. Laboratory Investigation, 93(4), 480-497. https://doi.org/10.1038/labinvest.2013.10