Methodological Aspects of Flow Cytometry DNA Analysis in Endometrial Carcinoma, with Special Reference to Sampling and Reproducibility

Thorkell E. Gudmundsson Department of Oncology, Division of Gynecologic Oncology (T.E. Gudmundsson, T. Högberg), and General Section (E. Långström B. Baldetorp, M. Fernö, D. Killander), University Hospital, Lund, Sweden; Department of Cytology and Pathology (P. Alm) and the Southern Swedish Regional Tumor Registry (H. Anderson), University Hospital, Lund, Sweden

Eva Långström Department of Oncology, Division of Gynecologic Oncology (T.E. Gudmundsson, T. Högberg), and General Section (E. Långström B. Baldetorp, M. Fernö, D. Killander), University Hospital, Lund, Sweden; Department of Cytology and Pathology (P. Alm) and the Southern Swedish Regional Tumor Registry (H. Anderson), University Hospital, Lund, Sweden

Per Alm Department of Oncology, Division of Gynecologic Oncology (T.E. Gudmundsson, T. Högberg), and General Section (E. Långström B. Baldetorp, M. Fernö, D. Killander), University Hospital, Lund, Sweden; Department of Cytology and Pathology (P. Alm) and the Southern Swedish Regional Tumor Registry (H. Anderson), University Hospital, Lund, Sweden

Harald Anderson Department of Oncology, Division of Gynecologic Oncology (T.E. Gudmundsson, T. Högberg), and General Section (E. Långström B. Baldetorp, M. Fernö, D. Killander), University Hospital, Lund, Sweden; Department of Cytology and Pathology (P. Alm) and the Southern Swedish Regional Tumor Registry (H. Anderson), University Hospital, Lund, Sweden

Bo Baldetorp Department of Oncology, Division of Gynecologic Oncology (T.E. Gudmundsson, T. Högberg), and General Section (E. Långström B. Baldetorp, M. Fernö, D. Killander), University Hospital, Lund, Sweden; Department of Cytology and Pathology (P. Alm) and the Southern Swedish Regional Tumor Registry (H. Anderson), University Hospital, Lund, Sweden

Mårten Fernö Department of Oncology, Division of Gynecologic Oncology (T.E. Gudmundsson, T. Högberg), and General Section (E. Långström B. Baldetorp, M. Fernö, D. Killander), University Hospital, Lund, Sweden; Department of Cytology and Pathology (P. Alm) and the Southern Swedish Regional Tumor Registry (H. Anderson), University Hospital, Lund, Sweden

Thomas Högberg Department of Oncology, Division of Gynecologic Oncology (T.E. Gudmundsson, T. Högberg), and General Section (E. Långström B. Baldetorp, M. Fernö, D. Killander), University Hospital, Lund, Sweden; Department of Cytology and Pathology (P. Alm) and the Southern Swedish Regional Tumor Registry (H. Anderson), University Hospital, Lund, Sweden

Dick Killander Department of Oncology, Division of Gynecologic Oncology (T.E. Gudmundsson, T. Högberg), and General Section (E. Långström B. Baldetorp, M. Fernö, D. Killander), University Hospital, Lund, Sweden; Department of Cytology and Pathology (P. Alm) and the Southern Swedish Regional Tumor Registry (H. Anderson), University Hospital, Lund, Sweden

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Abstract

Two adjacent paraffin-embedded sections manifested concordance in ploidy status in 96% of cases (45/47), and the standard deviation (SD) for SPF was 2.7%. Analysis of ‘micro-heterogeneity’, within a distance of ⩽700μ, yielded results for concordant ploidy status in 94% of cases, and the SD for SPF was 1.9% (n = 17). Frozen and paraffin-embedded material yielded concordant results for ploidy status in 87% (39/45) of cases, and SPF values were significantly lower (mean difference 1.5%) in the frozen samples. Diagnostic and repeat curettage material yielded concordant results for DNA ploidy status in 85% (40/47) of cases, and no significant difference in mean SPF (12% vs. 11%) was found. Discordant DNA ploidy results were attributable to small differences in the DNA histograms influencing the interpretation of near-diploid, near-tetraploid and small non-diploid cell populations, and the influence of debris on SPF estimation. On the basis of our findings and the practical advantage we recommend paraffin-embedded material from diagnostic curettage for FCM DNA analysis; the results are available sooner and the handling and transportation of tumor samples is more convenient.

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Methodological Aspects of Flow Cytometry DNA Analysis in Endometrial Carcinoma, with Special Reference to Sampling and Reproducibility

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Methodological Aspects of Flow Cytometry DNA Analysis in Endometrial Carcinoma, with Special Reference to Sampling and Reproducibility

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