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

How much formalin is enough to fix tissues?

Author(s): Buesa RJ, Peshkov MV

Publication: Ann Diagn Pathol, 2012, Vol. 16, Page 202-9

PubMed ID: 22483550 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to determine the effects of fixative volume to tissue ratio, tissue type, and fixation duration on sectioning quality, H and E staining, and vimentin immunostaining among formalin-fixed, paraffin-embedded (FFPE) tissue specimens.

Conclusion of Paper

Although not statistically significant, the average quality of sectioning performed by rotary microtome increased when tissue to fixative ratios decreased from 1:1 to 1:10. Significant increases in sectioning quality were observed when fixation time was increased from 8 h to 24 or 48 h. Among the 5 tissue types, fat and skin were the most difficult to section and were significantly harder than uterus, breast, and liver. Breast, uterus, and liver sectioning quality was not significantly different from one another. Neither H and E staining quality nor vimentin immunostaining (uterus only) were affected by fixation time or tissue to fixative ratio.

Studies

  1. Study Purpose

    The purpose of this study was to determine the effects of fixative volume to tissue ratio, tissue type, and fixation duration on sectioning quality, H and E staining, and vimentin immunostaining among FFPE tissue specimens. Uterus, breast, liver, abdominal skin, and underlying fat tissue were collected 36 h post-mortem after storage at 4-6 degrees C. FFPE specimens were sectioned using a rotary microtome.

    Summary of Findings:

    Although not statistically significant, the average sectioning quality (impossible scored as 0, bad scored as 1, fair scored as 2, good scored as 3) increased when tissue to fixative ratios decreased from 1:1 to 1:10. Significant increases in sectioning quality were observed when fixation time was increased from 8 h to 24 or 48 h. Among the 5 tissue types, fat and skin were the most difficult to section and were significantly harder than uterus, breast, and liver. Breast, uterus, and liver sectioning quality was not significantly different from one another. Neither H and E staining quality nor vimentin immunostaining (uterus only) were affected by fixation time or tissue to fixative ratio.

    Biospecimens
    Preservative Types
    • Formalin
    Diagnoses:
    • Autopsy
    • Normal
    Platform:
    AnalyteTechnology Platform
    Morphology H-and-E microscopy
    Morphology Macroscopic observation
    Protein Immunohistochemistry
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Biospecimen Acquisition Biospecimen location Uterus
    Breast
    Liver
    Abdominal skin
    Abdominal fat
    Biospecimen Preservation Time in fixative 8 h
    24 h
    48 h
    Biospecimen Preservation Tissue to fixative ratio 1:1
    1:2
    1:5
    1:10
    Immunohistochemistry Specific Targeted peptide/protein Vimentin

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