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Related Experiment Videos

Hematoxylin staining in quantitative DNA cytometry: an image analysis study

E K Schulte1, D K Fink

  • 1Anatomisches Institut, Johannes Gutenberg-Universität, Mainz, Germany.

Analytical Cellular Pathology : the Journal of the European Society for Analytical Cellular Pathology
|December 1, 1995
PubMed
Summary

Aluminum-hematein is unsuitable for quantitative DNA image cytometry. Studies showed it does not stain DNA stoichiometrically, leading to significant photometric errors, making it unreliable for accurate DNA content analysis.

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Area of Science:

  • Biomedical Imaging
  • Cytometry
  • Histopathology

Background:

  • Accurate quantification of DNA content is crucial in diagnostics.
  • Aluminum-hematein is a common histological stain, but its quantitative DNA analysis capabilities are not well-established.

Purpose of the Study:

  • To evaluate the suitability of aluminum-hematein for quantitative DNA image cytometry.
  • To compare pure and commercial aluminum-hematein formulations.

Main Methods:

  • Staining of cervical smears, breast cancer biopsies, and rabbit liver imprints with Mayer's and Harris' aluminum-hematein.
  • Nucleic acid removal via enzyme digestion or HCl-hydrolysis.
  • Use of DNA-polyacrylamide films as staining models.
  • Absorption measurement using a VIDAS image analyzer.

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  • Comparison with Feulgen stain as a control.

    • Main Results:

    • Aluminum-hematein did not demonstrate a stoichiometric dye-DNA ratio in liver cell nuclei.
    • Sequential staining with hematein and Feulgen reaction showed moderate covariance (0.77-0.88) for integrated optical density (IOD).
    • Significant photometric errors resulted from non-specific RNA and protein staining.
    • Pure hematein yielded slightly better, though still non-quantitative, results compared to commercial batches.
    • DNA staining in model films was not quantitative with hematein.

    Conclusions:

    • Aluminum-hematein is not recommended for quantitative DNA image cytometry due to lack of stoichiometry and significant error sources.
    • The stain's unreliability for DNA quantification necessitates alternative methods for accurate analysis.