NIH, National Cancer Institute, Division of Cancer Treatment and Diagnosis (DCTD) NIH - National Institutes of Health National Cancer Institute DCTD - Division of Cancer Treatment and Diagnosis

Optimal tumor sampling for immunostaining of biomarkers in breast carcinoma.

Author(s): Tolles J, Bai Y, Baquero M, Harris LN, Rimm DL, Molinaro AM

Publication: Breast Cancer Res, 2011, Vol. 13, Page R51

PubMed ID: 21592345 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to determine the effects of the number of images analyzed using the Automated Quantitative Analysis (AQUA) method on the quantification of immunofluorescence for several cancer biomarkers.

Conclusion of Paper

The optimum number of 20x field images required to estimate the expression of a biomarker in a whole tissue section, as assessed by AQUA score, varied based on biomarker and ranged from 3 to 14.

Studies

  1. Study Purpose

    The purpose of this study was to determine the effects of the number of 20x field images analyzed using AQUA on the quantification of immunofluorescence for cancer biomarkers including estrogen receptor (ER), human epidermal growth factor receptor 2 (HER2), serine/threonine protein kinase (AKT), extracellular-signal-regulated kinase (ERK), ribosomal protein S6 kinase beta-1 (S6K1), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), cytokeratin, and microtubule-associated protein (MAP)-Tau. Due to the small sample size and number of fields of view, a mathematical simulation was performed for all biomarkers except MAP-Tau, for which direct sampling of 122 whole section slides and the simulation were performed. Only ER-positive breast cancer cases were included.

    Summary of Findings:

    The optimum number of 20x field images required to estimate the expression of a biomarker in a whole tissue section, as assessed by AQUA score, varied based on biomarker and ranged from 3 (cytokeratin) to 14 (MAP-Tau, simulated and direct sampling). In general, a higher number of 20x fields of view were required for markers showing more heterogeneity.

    Biospecimens
    Preservative Types
    • Formalin
    Diagnoses:
    • Neoplastic - Carcinoma
    Platform:
    AnalyteTechnology Platform
    Protein Immunohistochemistry
    Morphology Fluorescent microscopy
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Biospecimen Aliquots and Components Aliquot size/volume 1-35 20x fields of view
    Immunohistochemistry Specific Targeted peptide/protein ER
    HER2
    AKT
    ERK
    S6K1
    GAPDH
    Cytokeratin
    MAP-Tau

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