Diff-Quik: A reliable stain for rapid onsite evaluation and differentiation of round cells in semen

Abstract

Objectives: Identification of immature germ cells and leukocytes and their differentiation pose difficulty in interpretation. In this study, Diff-Quik (DQ) and Papanicolaou (Pap) are used to differentiate between these two, as they are recommended by the WHO, cost-effective and are easy to use. Materials and Methods: This study attempts to differentiate between immature germ cells and leukocytes in semen; compare the usefulness of two stains –Pap and DQ; evaluate the possible associations between the total sperm count and the round cell count; the total sperm count and the presence of round cells and inflammatory cells; the occupation and the sperm concentration; and the occupation and the round cell count. Results: A total of 60 semen samples were included in the study. Two smears were made for each patient; DQ stain was used to stain the air-dried smears and Pap stain for alcohol-fixed smears. The whole procedure of DQ staining takes only 30 s. The smears were compared. There was good concordance between DQ and Pap stains in the detection of inflammatory cells and immature germ cells (P = 0.000). There was no association between total sperm count and round cell count (r = −0.086 and P = 0.520); and between the occupation and the sperm concentration or the round cell count. Conclusion: There was an inverse relationship between total sperm count and presence of immature germ cells (r = −0.291). There was no association between round cell count and presence of immature germ cells (r = 0.148 and P = 0.269), total sperm count and presence of inflammatory cells, round cell count, and presence of inflammatory cells. DQ stain is easy to use, convenient, and rapid. There is good concordance between DQ and Pap stains in the detection of inflammatory cells and immature germ cells. Total sperm count is independent of the round cell count or the presence of inflammatory cells/immature germ cells. In most of the cases, the round cells were the inflammatory cells. Morphological interpretation can be done even in non-liquefied samples.