ABSTRACT
PURPOSE: We evaluated the validity and reproducibility of sperm morphology analysis using strict criteria to compare the difference in the percentages of normal and amorphous morphology as seen by two staining methods (Diff-Quik and Papanicolaou)with the eventual goal of developing a computer-assisted sperm morphology assessment system. MATERIALS AND METHODS: Semen samples were obtained from 10 volunteers and stained by the Diff-Quik and Papanicolaou staining methods. Under 1000-fold oil-immersion magnification, the slides were examined by two experienced observers, and the screens were transferred to the monitor using a CCD camera. A calibrated transparent overlay was used to distinguish between normal and abnormal morphology according to strict criteria. One hundred spermatozoa for each sample were analyzed by two experienced observers. RESULTS: In Diff-Quik staining, intraobserver and interobserver correlation coefficients for normal and amorphous sperm were 0.862 (p<0.05), 0.916 (p<0.05) and 0.857 (p<0.05), 0.890 (p<0.05), respectively. In Papanicolaou staining, intraobserver correlation coefficients for normal and amorphous sperm were 0.832 (p<.05), 0.877 (p<0.05) and 0.846 (p<0.05) and 0.865 (p<0.05), respectively. CONCLUSIONS: Assessing normal and abnormal spem abnormal sperm morphology using strict criteria produces reproducible results between and within observers. Also, there were good correlations between the two staining methods in assessing normal and amorphous morphology. This study presents the basic data for the development of computer-assisted sperm morphology assessment system. New techniques should be developed to analyze amorphous sperm.
Subject(s)
Semen , Spermatozoa , VolunteersABSTRACT
PURPOSE: We evaluated the validity and reproducibility of sperm morphology analysis using strict criteria to compare the difference in the percentages of normal and amorphous morphology as seen by two staining methods (Diff-Quik and Papanicolaou)with the eventual goal of developing a computer-assisted sperm morphology assessment system. MATERIALS AND METHODS: Semen samples were obtained from 10 volunteers and stained by the Diff-Quik and Papanicolaou staining methods. Under 1000-fold oil-immersion magnification, the slides were examined by two experienced observers, and the screens were transferred to the monitor using a CCD camera. A calibrated transparent overlay was used to distinguish between normal and abnormal morphology according to strict criteria. One hundred spermatozoa for each sample were analyzed by two experienced observers. RESULTS: In Diff-Quik staining, intraobserver and interobserver correlation coefficients for normal and amorphous sperm were 0.862 (p<0.05), 0.916 (p<0.05) and 0.857 (p<0.05), 0.890 (p<0.05), respectively. In Papanicolaou staining, intraobserver correlation coefficients for normal and amorphous sperm were 0.832 (p<.05), 0.877 (p<0.05) and 0.846 (p<0.05) and 0.865 (p<0.05), respectively. CONCLUSIONS: Assessing normal and abnormal spem abnormal sperm morphology using strict criteria produces reproducible results between and within observers. Also, there were good correlations between the two staining methods in assessing normal and amorphous morphology. This study presents the basic data for the development of computer-assisted sperm morphology assessment system. New techniques should be developed to analyze amorphous sperm.