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 language | English (US) |
|---|---|
| Pages (from-to) | 480-497 |
| Number of pages | 18 |
| Journal | Laboratory Investigation |
| Volume | 93 |
| Issue number | 4 |
| DOIs | |
| State | Published - Apr 2013 |
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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
Cite this
Quantitation of fixative-induced morphologic and antigenic variation in mouse and human breast cancers. / Cardiff, Robert; Hubbard, Neil; Engelberg, Jesse A.; Munn, Robert J.; Miller, Claramae H.; Walls, Judith E.; Chen, Jane Q.; Velásquez-García, Héctor A.; Galvez, Jose J.; Bell, Katie J.; Beckett, Laurel A; Li, Yue Ju; Borowsky, Alexander D.
In: Laboratory Investigation, Vol. 93, No. 4, 04.2013, p. 480-497.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Quantitation of fixative-induced morphologic and antigenic variation in mouse and human breast cancers
AU - Cardiff, Robert
AU - Hubbard, Neil
AU - Engelberg, Jesse A.
AU - Munn, Robert J.
AU - Miller, Claramae H.
AU - Walls, Judith E.
AU - Chen, Jane Q.
AU - Velásquez-García, Héctor A.
AU - Galvez, Jose J.
AU - Bell, Katie J.
AU - Beckett, Laurel A
AU - Li, Yue Ju
AU - Borowsky, Alexander D
PY - 2013/4
Y1 - 2013/4
N2 - 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.
AB - 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.
KW - estrogen receptor
KW - Her2
KW - IHC4 biomarker panel
KW - immunohistochemistry
KW - MIB1 (Ki67)
KW - progesterone receptor
KW - quantitative image analysis
UR - http://www.scopus.com/inward/record.url?scp=84875707939&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84875707939&partnerID=8YFLogxK
U2 - 10.1038/labinvest.2013.10
DO - 10.1038/labinvest.2013.10
M3 - Article
VL - 93
SP - 480
EP - 497
JO - Laboratory Investigation
JF - Laboratory Investigation
SN - 0023-6837
IS - 4
ER -