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Establishment of a diploid reference value for DNA ploidy analysis by image cytometry in mouse cells
M dos Anjos Pires1, C Palmeira, P Rodrigues
1Institute for Agrarians and Agro-Food Sciences and Technologies, Laboratory of Histology and Anatomic Pathology, Universidade De Trás-os-Montes e Alto Douro, Vila Real, Portugal. apires@utad.pt
Objective:
To establish a diploid reference value for DNA ploidy analysis of mouse cells (Mus musculus) by image cytometry using the CAS 200, an analysis system suitable for DNA content studies in human cells.
Study Design:
To establish this standard, we used spleen imprints from 26 normal animals. A minimum of 150 lymphocytes present in each imprint was counted. The mean DNA content (pg/cell) of the G0/G1 peak and the DNA index observed in all samples were statistically analyzed. Cytospins with peritoneal cells from the same animals were then analyzed with this reference DNA value to confirm the diploid range.
Results:
The DNA diploid reference value was determined by the mean DNA content of all spleen samples, which was 6.42 +/- 0.234 pg/cell, and the diploid range, defined as the diploid value +/- 10%, was 5.78-7.06 pg/cell. All the peritoneal samples showed a DNA diploid histogram, with a mean value for the G0/G1 peak DNA content of 6.742 +/- 0.15.
Conclusion:
The diploid reference value found in this study differs from those reported for other species, including the human being, and should be used in further studies of mouse pathology.