Histological Techniques for Mammary Tissue Analysis

Tissues were fixed in 10% neutral-buffered formalin, and processed and embedded according to standard histologic protocols. Five-micron sections were stained with hematoxylin and eosin (H&E) or used for immunohistochemical analysis. The GLUT1 antibody has been described previously [31 (link)]. Anti-Cre recombinase antibody was from Thermo Scientific, anti-Ki67 antibody was obtained from Dako, and anti-BrdU antibody was from Abcam. Cre recombinase and Ki67 were quantified by counting the number of positive cells out of the total number of mammary epithelial cells in one full, representative mammary gland section per mouse. BrdU staining was quantified by counting the number of positive cells out of the total cells per acinar structure in 3–4 paraffin-embedded sections per treatment, cut every 50 μm. GLUT1 and tumor area were quantified using the Aperio Digital Pathology system (Leica Biosystems).

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Wellberg E.A., Johnson S., Finlay-Schultz J., Lewis A.S., Terrell K.L., Sartorius C.A., Abel E.D., Muller W.J, & Anderson S.M. (2016). The glucose transporter GLUT1 is required for ErbB2-induced mammary tumorigenesis. Breast Cancer Research : BCR, 18, 131.

Publication 2016

anti-Ki67 antibody anti-BrdU antibody Aperio Digital Pathology system

Acinar cells structure Anti antibody Antibody Brdu Cre recombinase Embedded paraffin Eosin Formalin Glut1 Mammary Mammary gland Mouse Tissues Tumor

Corresponding Organization : University of Colorado Anschutz Medical Campus

Other organizations : University of Iowa, McGill University

Top 5 similar protocols

Variable analysis

independent variables

Antibody used for immunohistochemical analysis (GLUT1, Cre recombinase, Ki67, BrdU)

dependent variables

GLUT1 expression
Cre recombinase positive cells
Ki67 positive cells
BrdU positive cells
Tumor area

control variables

Tissue processing and embedding according to standard histologic protocols
Hematoxylin and eosin (H&E) staining

positive controls

GLUT1 antibody previously described [31]

negative controls

None specified

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