Use of cultured cells as a control for quantitative immunocytochemical analysis of estrogen receptor in breast cancer: The Quicgel method

• Published in: American journal of clinical pathology Vol. 111; no. 3; pp. 329 - 335
• Main Authors: RIERA, J, SIMPSON, J. F, TAMAYO, R, BATTIFORA, H, MD
• Format: Journal Article
• Language: English
• Published: Chicago, IL American Society of Clinical Pathologists 01.03.1999
• Subjects: Adult; Aged; Aged, 80 and over; Biological and medical sciences; Breast Neoplasms - chemistry; Breast Neoplasms - pathology; Carcinoma - chemistry; Carcinoma - pathology; Cell Count; Female; Humans; Image Processing, Computer-Assisted; Immunoenzyme Techniques; Investigative techniques, diagnostic techniques (general aspects); Medical sciences; Middle Aged; Miscellaneous. Technology; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques; Quality Control; Receptors, Estrogen - analysis; Reproducibility of Results; Tumor Cells, Cultured - chemistry; Tumor Cells, Cultured - pathology; Human; Immunohistochemistry; Cell culture; Carcinoma; Estrogen; Malignant tumor; Ovarian hormone; Pathology; Mammary gland diseases; Quality control; Established cell line; Mammary gland; Technique; Sex steroid hormone; Freeze; Quantitative analysis; Hormonal receptor
• ISSN: 0002-9173; 1943-7722
• DOI: 10.1093/ajcp/111.3.329

Variation in tissue fixation, processing, and staining is largely responsible for poor reproducibility of estrogen receptor (ER) immunohistochemistry assays. A frozen, agar-suspended pellet of MCF-7 cells with known ER content was added to each of 55 samples of invasive breast carcinoma (IBC), serving as a control. Image analysis determined percentages of positive area (positive nuclei per total nuclei analyzed) and positive stain (sum of optical density of the positive nuclear area divided by sum of the optical density of all nuclei studied) of MCF-7 cells and IBC. MCF-7 cells had a mean value of 150 fmol/mg of ER by dextran-coated charcoal analysis. Image analysis of MCF-7 cells included with the 55 cases showed a mean positive area of 70.81. Positive staining from the IBC cases ranged from 0 to 98.5. By using the known ER content and the positive area of the MCF-7 cells, a conversion factor was used to translate the positive area of the clinical specimens to a femtomole equivalent, which for the 55 IBCs ranged from 0 to 1,790 (mean, 187). Inclusion of a control with known femtomole quantity of ER provides an internal standard for quality control and ER quantitation.