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

Rapid microwave-stimulated fixation of entire prostatectomy specimens. Biomed-II MPC Study Group.

Author(s): Ruijter ET, Miller GJ, Aalders TW, van de Kaa CA, Schalken JA, Debruyne FM, Boon ME

Publication: J Pathol, 1997, Vol. 183, Page 369-75

PubMed ID: 9422995 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to determine the effects of method of fixative delivery, time in fixative and antigen retrieval on morphology, protein and DNA detection.

Conclusion of Paper

The use of formalin perfusion followed by two rounds of microwave irradiation allowed for homogeneous histological and immunohistochemical (IHC) staining and extraction of large DNA fragments. In contrast, immersion fixation resulted in edge effects for some IHC targets and H&E staining as well as DNA that was mostly under 1 kb in length. When immersion fixation was used, optimal fixation time for E-cadherin staining was 8-12 h, but p53 and MIB-1 staining did not depend on fixation duration or method of delivery. Cytokeratin staining in immersion fixed tissue was optimal when fixation was for 12 h or less and when protease digestion was used.

Studies

  1. Study Purpose

    The purpose of this study was to determine the effects of method of fixative delivery on the histology of prostate specimens obtained at autopsy.

    Summary of Findings:

    Sectioning of specimens which were fixed through immersion in neutral buffered formalin or microwave-accelerated fixation for 6 min followed by 3.5 min was easy, regardless of whether the tissue was previously perfused through multiple injections of formalin. However, if the tissue underwent microwave-accelerated fixation for only 6 min, with or without perfusion, sectioning was difficult and resulted in damage to the tissue. Tissue that was not perfused with formalin prior to H&E staining displayed an edge effect. The authors report that there were no significant differences between conventional immersion and microwave-accelerated fixation for PAS, van Gieson's elastica or Alcian blue staining.

    Biospecimens
    Preservative Types
    • Formalin
    Diagnoses:
    • Autopsy
    • Not specified
    Platform:
    AnalyteTechnology Platform
    Morphology H-and-E microscopy
    Morphology Light microscopy
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Biospecimen Preservation Method of fixative delivery Immersion
    Microwaved
    Injection
  2. Study Purpose

    The purpose of this study was to determine the effects of method of fixative delivery on immunohistochemistry (IHC) of prostate specimens obtained at autopsy.

    Summary of Findings:

    E-cadherin staining required antigen retrieval and was most intense when conventional immersion fixation was for 8-12 h. When conventional immersion fixation was used, most E-cadherin staining was peripheral or absent with only 16% of specimens displaying specific homogeneous staining. Multiple perfusion injections prior to immersion fixation increased the number of specimens with specific homogeneous E-cadherin staining to 53%. When perfusion was followed by two rounds of microwave irradiation, 93% of the specimens had specific homogeneous E-cadherin staining. Method of fixative delivery and duration of fixation had no effects on p53 or MIB-1 staining. Cytokeratin staining in conventionally fixed specimens decreased slightly when fixation was for more than 12 h, but was increased by protease digestion. Cytokeratin staining was homogeneous in all specimens fixed by perfusion followed by two rounds of microwave irradiation.

    Biospecimens
    Preservative Types
    • Formalin
    Diagnoses:
    • Autopsy
    • Not specified
    Platform:
    AnalyteTechnology Platform
    Protein Immunohistochemistry
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Biospecimen Preservation Method of fixative delivery Immersion
    Injection
    Microwaved
    Biospecimen Preservation Time in fixative 1 h
    2 h
    3 h
    4 h
    8 h
    12 h
    18 h
    24 h
    36 h
    Immunohistochemistry Specific Targeted peptide/protein Cytokeratin
    E-cadherin
    MIB-1
    p53
    Analyte Extraction and Purification Antigen retrieval Microwave in 0.1 M citrate buffer
    None
    Analyte Extraction and Purification Protein digestion 10 min digestion in Protease XIV at 37 degrees C
    None
  3. Study Purpose

    The purpose of this study was to determine the effects of method of fixative delivery on the size of extracted DNA from prostate specimens obtained at autopsy.

    Summary of Findings:

    When specimens were fixed by immersion in formalin for 20 h, only 34% of the DNA was over 1 kb and 2-3% was over 4 kb. In contrast, when the method of fixative delivery included multiple perfusion injections followed by two rounds of microwave irradiation, 78% of the extracted DNA was over 1 kb and 40% was over 4 kb.

    Biospecimens
    Preservative Types
    • Formalin
    Diagnoses:
    • Autopsy
    • Not specified
    Platform:
    AnalyteTechnology Platform
    DNA Electrophoresis
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Biospecimen Preservation Method of fixative delivery Immersion
    Microwaved
    Injection

You Recently Viewed  

News and Announcements

  • Most Downloaded SOPs in 2024

  • New Articles on the GTEx Project are Now FREELY Available!

  • Just Published!

  • More...