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Related Experiment Videos

Variations associated with disaggregation methods in DNA flow cytometry

T K Lee1, A L Wiley, J D Esinhart

  • 1Department of Radiation Oncology, Leo W. Jenkins Cancer Center, East Carolina University School of Medicine, Greenville, North Carolina 27858.

Analytical and Quantitative Cytology and Histology
|June 1, 1993
PubMed
Summary
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Scraping tumor tissue yields the most DNA nondiploid clones for flow cytometry (FC) analysis. Multiple sampling methods provide comprehensive DNA ploidy information for solid tumors.

Area of Science:

  • Oncology
  • Biotechnology
  • Cell Biology

Background:

  • Accurate DNA ploidy analysis is crucial for solid tumor characterization.
  • Different tissue disaggregation methods can impact cell suspension quality and subsequent flow cytometry (FC) results.
  • Variations in cell yield, viability, and malignant cell percentage may affect DNA ploidy assessment.

Purpose of the Study:

  • To investigate the influence of various disaggregation methods on DNA ploidy analysis by flow cytometry (FC).
  • To determine the optimal method for obtaining cell suspensions for reliable DNA ploidy assessment in solid tumors.

Main Methods:

  • Solid tumor specimens (n=40) were disaggregated using mechanical mincing, enzymatic digestion, in vitro fine needle aspiration (FNA), and scraping.
  • Cell suspensions were analyzed for cell yield, viability, and DNA ploidy using flow cytometry (FC).

Related Experiment Videos

  • DNA ploidy concordance was assessed across different disaggregation methods for the same tumor specimens.

    • Main Results:

    • Mechanical mincing yielded the highest cell count, while enzymatic digestion offered the best cell viability.
    • No significant differences were observed in G0/G1 coefficient of variation, DNA indices, or S phase percentage across methods.
    • Scraping method resulted in the highest percentage of malignant cells (82.3%) and the highest detection rate of DNA nondiploid clones (61.3%).

    Conclusions:

    • Scraping tumor tissue is the preferred method for DNA ploidy analysis by FC due to its higher yield of nondiploid clones.
    • Employing multiple disaggregation techniques enhances the comprehensive assessment of DNA ploidy in solid tumors.
    • Standardizing disaggregation methods is essential for reproducible and accurate FC-based tumor ploidy studies.