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

A novel fixative improves opportunities of nucleic acids and proteomic analysis in human archive's tissues.

Author(s): Stanta G, Mucelli SP, Petrera F, Bonin S, Bussolati G

Publication: Diagn Mol Pathol, 2006, Vol. 15, Page 115-23

PubMed ID: 16778593 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to compare the preservation of nucleic acids and proteins in tissue preserved with FineFIX or formalin and embedded in paraffin as well as with fresh frozen tissue.

Conclusion of Paper

The authors report that tissues preserved in FineFIX resulted in more intense immunohistochemical staining without compromising morphology, greater DNA yield, and greater DNA and RNA integrity that produced longer PCR generated amplicons compared to formalin-fixed specimens. Further, the authors noted successful analysis of protein extracts with respect to pattern and expression in FineFIXed specimens, yielding comparable results to fresh frozen controls.

Studies

  1. Study Purpose

    The purpose of this study was to compare protein antigenicity via immunohistochemistry among tissues preserved with FineFIX or formalin and paraffin embedded.

    Summary of Findings:

    Compared to formalin-fixed, paraffin-embedded tissue, tissue fixed in FineFIX with microwave assistance and embedded in paraffin yielded greater staining intensity while preserving appropriate staining patterns.

    Biospecimens
    Preservative Types
    • Other Preservative
    • Formalin
    Diagnoses:
    • Neoplastic - Carcinoma
    Platform:
    AnalyteTechnology Platform
    Protein Immunohistochemistry
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Biospecimen Preservation Type of fixation/preservation FineFIX
    Formalin (buffered)
    Biospecimen Preservation Method of fixative delivery Immersion
    Microwaved
  2. Study Purpose

    The purpose of this study was to compare RNA integrity in FineFIXed and formalin-fixed specimens that were paraffin embedded.

    Summary of Findings:

    Longer beta-actin amplicons were generated from RNA extracted from FineFixed, paraffin-embedded specimens (450 bp in 100% and 600 bp in 60% of cases) compared to RNA extracted from formalin-fixed, paraffin embedded specimens (170 bp).

    Biospecimens
    Preservative Types
    • Other Preservative
    • Formalin
    Diagnoses:
    • Neoplastic - Carcinoma
    Platform:
    AnalyteTechnology Platform
    RNA RT-PCR
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Biospecimen Preservation Type of fixation/preservation FineFIX
    Formalin (buffered)
    RT-PCR Specific Length of gene fragment 77 bp
    170 bp
    200 bp
    438 bp
    651 bp
  3. Study Purpose

    The purpose of this study was to compare the preservation of DNA in FineFIXed and formalin-fixed specimens that were paraffin embedded.

    Summary of Findings:

    The authors reported greater total DNA yield from FineFIXed specimens compared to those fixed in formalin. The maximum TTR fragment length amplified from DNA isolated from formalin-fixed, paraffin-embedded tissue sections was 339bp, while tissue fixed in FineFIX and embedded paraffin yielded fragments as long as 2395 bp, the longest amplicon attempted.

    Biospecimens
    Preservative Types
    • Other Preservative
    • Formalin
    Diagnoses:
    • Neoplastic - Carcinoma
    Platform:
    AnalyteTechnology Platform
    DNA PCR
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Biospecimen Preservation Type of fixation/preservation FineFIX
    Formalin (buffered)
    PCR Specific Length of gene fragment 291 bp
    339 bp
    1000 bp
    1939 bp
    2395 bp
  4. Study Purpose

    The purpose of this study was to determine if protein extraction and subsequent analysis was possible in FineFIXed specimens, and whether results were comparable to those observed in fresh frozen specimens.

    Summary of Findings:

    Of the approximately 1000 protein spots identified by 2D-gel analysis, only 1.4% differed among FineFIXed and fresh frozen specimens. Western blot analysis for alpha tubulin and human tTG yielded comparable results among FineFIXed and fresh frozen specimens, although a second smaller alpha tubulin protein band was observed in fresh frozen specimens, which the authors hypothesized to be a product of degradation. No proteins were observed in extracts from formalin-fixed specimens.

    Biospecimens
    Preservative Types
    • Other Preservative
    • Frozen
    • Formalin
    Diagnoses:
    • Neoplastic - Carcinoma
    Platform:
    AnalyteTechnology Platform
    Protein 1D/2D gels
    Protein Western blot
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Biospecimen Preservation Type of fixation/preservation FineFIX
    Formalin (buffered)
    Snap frozen

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