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
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An immunohistochemical evaluation of progesterone receptor in frozen sections, paraffin sections, and cytologic imprints of breast carcinomas.

Author(s): Ozzello L, DeRosa C, Habif DV, Greene GL

Publication: Cancer, 1991, Vol. 67, Page 455-62

PubMed ID: 1845948 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to determine the effects of different preservation methods, antibody types, and technology platforms on progesterone receptor (PR) and estrogen receptor (ER) detection using frozen specimens and fixed, paraffin embedded specimens as well as stored cryostat sections and imprints of breast cancer specimens.

Conclusion of Paper

ER immunohistochemical (IHC) staining of paraffin-embedded specimens gave the best results with fixations of 1-2 hours in formalin at 4 degrees C or in Bouin's solution at room temperature. Concordance rates between PR and ER immunostaining of frozen and paraffin sections were 85.4 and 93.6%, respectively. Concordance rates between PR and ER immunostaining of specimens stored for up to 56 weeks and frozen sections were each 92.9%. No differences were seen between the antibodies used for IHC staining. Overall, more ER+ and/or PR+ cases were identified by IHC staining than by the dextran-coated charcoal assay (DCCA). Concordance rates between the two assays ranged from 77.2 to 80.8% for PR and from 78.6 to 91.4% for ER.

Studies

  1. Study Purpose

    The purpose of this study was to determine the effects of fixative, and duration and temperature of fixation on ER and PR immunostaining results using frozen and fixed paraffin embedded specimens as well as stored cryostat sections and imprints of breast cancer specimens.

    Summary of Findings:

    ER IHC staining of paraffin-embedded specimens yielded the best results with short fixations of 1-2 hours in formalin at 4 degrees C or in Bouin's solution at room temperature. A fixation time of 24 hours in formalin or Bouin's resulted in some nonspecific cytoplasmic staining, except in formalin at 4 degrees C. PR immunostaining was good in imprints and cryostat sections stored at -80 degrees C when they were fixed in Zamboni's before freezing. Concordance rates between PR and ER immunostaining of frozen and paraffin sections were 85.4 and 93.6%, respectively. The authors said it was difficult to determine which preservation technique gave the most accurate results. Concordance rates between PR and ER immunostaining of preparations stored for up to 56 weeks and frozen sections were each 92.9%. Coating of imprints and cryostat sections with 6% polyethylene glycol during storage impaired immunoreactivity.

    Biospecimens
    Preservative Types
    • Formalin
    • Other Preservative
    • Frozen
    Diagnoses:
    • Neoplastic - Carcinoma
    Platform:
    AnalyteTechnology Platform
    Protein Immunohistochemistry
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Biospecimen Preservation Type of fixation/preservation Bouin's fixative
    Formalin (buffered)
    Frozen
    Zamboni's solution
    Immunohistochemistry Specific Targeted peptide/protein ER
    PR
    Biospecimen Aliquots and Components Cell capture method Cryostat section
    Imprint
    Paraffin section
    Biospecimen Preservation Temperature of fixation/preservation Room temperature
    4 degrees C
    Biospecimen Preservation Time in fixative 10 min
    30 min
    1 h
    2 h
    4 h
    24 h
    Storage Storage duration None
    Up to 56 weeks
    Storage Storage conditions Coated with 6% polyethylene glycol
    Uncoated
    View more
  2. Study Purpose

    The purpose of this study was to determine the effects of antibody type and to compare immunoassay results with IHC staining results for differently preserved breast cancer specimens.

    Summary of Findings:

    When used at different optimal concentrations, PR antibodies JZB39 and KD68 worked well for fresh or stored frozen sections, paraffin sections, and imprints, regardless of preservation method, and gave concordant results 96.1% of the time. Immunostaining heterogeneity was seen for both antibodies in all specimens. Overall, more ER+ and/or PR+ cases were identified by IHC staining than by the DCCA for both pre- and postmenopausal women. Concordance rates for PR results between the two assays for frozen sections, paraffin sections, and stored preparations were 77.2, 80.8, and 76.8%, respectively (p<0.0005). Concordance rates for ER results between the two assays for frozen sections, paraffin sections, and stored preparations were 91.4, 87.3, and 78.6%, respectively.

    Biospecimens
    Preservative Types
    • Formalin
    • Other Preservative
    • Frozen
    Diagnoses:
    • Neoplastic - Carcinoma
    Platform:
    AnalyteTechnology Platform
    Protein Immunohistochemistry
    Protein Immunoassay
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Immunohistochemistry Specific Targeted peptide/protein ER
    PR
    Immunohistochemistry Specific Type of antibody JZB39
    KD68
    H222
    D75P3gamma
    Immunoassay Specific Targeted peptide/protein ER
    PR
    Biospecimen Preservation Type of fixation/preservation Bouin's fixative
    Formalin (buffered)
    Frozen
    Zamboni's solution
    Biospecimen Aliquots and Components Cell capture method Cryostat section
    Imprint
    Paraffin section
    Immunohistochemistry Specific Technology platform Immunoassay
    Preaquisition Patient age Premenopausal
    Postmenopausal
    View more

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