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Reproducibility of DNA ploidy and S-phase values from paraffin-embedded tissue
L J Elavathil1, G Celebre, D McFarlane
1Department of Pathology, McMaster University Medical Centre, Ontario, Canada.
Objective:
To determine the degree of variability of DNA flow cytometric results between two types of flow cytometers.
Study Design:
Single-cell suspensions were made from 40 paraffin blocks of ovarian carcinoma using a modified Hedley procedure. These samples were consecutively analyzed in both Coulter's Profile II and Elite Flow cytometers. Gated and ungated data were collected and analyzed using Phoenix flow system's multicycle software.
Results:
There were 16 (40%) DNA diploid, 23 (58%) DNA aneuploid and 1 DNA tetraploid tumor. The degree of variability in the DNA index coefficient of variation of the G0/G1 diploid peak, percentage of S phase, percentage of G2M, percentage of debris plus percentage of clumps were compared. Excellent correlations of the results were obtained in the DNA index (r = .999) and in percentage of S phase (r = .946).
Conclusion:
It is feasible to standardize variables of flow cytometric instruments to obtain reproducible results.