Computer‐assisted analysis of sperm morphology with the aid of lectin staining

Summary.  Lectins are useful for staining the acrosome, which is a pre‐requisite for the assessment of acrosome reaction in vitro. We tested wheat germ agglutinin, peanut agglutinin and pisum sativum agglutinin. The determination of the categories of normal and abnormal spermatozoa as defined by the World Health Organization, including an additional category ’acrosome‐less cell’, was performed with the aid of a system of computer‐assisted semen analysis (CASA) in comparison with visual estimation. The acrosome reaction in vitro was induced by calcium ionophore A23187. Incubation with A23187 decreased the percentage of normal sperm heads and increased the number of acrosome‐less sperm heads in comparison with the control samples. This shift was demonstrable with all three lectin staining procedures. No significant differences were observed in the comparison of sperm classes obtained by visual assessment or determination by CASA with two of the lectin staining procedures. After staining with pisum sativum agglutinin the classes of normal and of acrosome‐less spermatozoa were significantly different between visual and CASA estimation. Our results indicate that estimation of the acrosomal status in vitro is possible by CASA when lectin staining is used.

[1]  W. Voorhout,et al.  Use of lectins to characterize plasma membrane preparations from boar spermatozoa: a novel technique for monitoring membrane purity and quantity. , 1998, Biology of reproduction.

[2]  C. Lombard,et al.  A new computerized method of reading sperm morphology (strict criteria) is as efficient as technician reading. , 1993, Fertility and sterility.

[3]  R. Menkveld,et al.  Comparison of normal sperm morphology outcomes from two different computer‐assisted semen analysis systems , 2001, Andrologia.

[4]  S. Nozawa,et al.  Quantitative assessment of human sperm acrosome reaction by using fluorescein isothiocyanate conjugated concanavalin A--comparison between highly purified acrosome reacted with non-acrosome reacted sperm. , 1998, Archives of andrology.

[5]  D. Liu,et al.  Morphology of spermatozoa bound to the zona pellucida of human oocytes that failed to fertilize in vitro. , 1992, Journal of reproduction and fertility.

[6]  T. Cooper,et al.  A flow cytometric technique using peanut agglutinin for evaluating acrosomal loss from human spermatozoa. , 1998, Journal of andrology.

[7]  W. Voorhout,et al.  Use of peanut agglutinin to assess the acrosomal status and the zona pellucida-induced acrosome reaction in stallion spermatozoa. , 1996, Journal of andrology.

[8]  J. Tesarik,et al.  Distinction between true acrosome reaction and degenerative acrosome loss by a one‐step staining method using Pisum sativum agglutinin , 1993, Journal of reproduction and fertility.

[9]  C. Matás,et al.  Lectin histochemistry during in vitro capacitation and acrosome reaction in boar spermatozoa: new lectins for evaluating acrosomal status of boar spermatozoa. , 1996, Acta histochemica.

[10]  G Richoilley,et al.  Computer-assisted assessment of sperm morphology: comparison with conventional techniques. , 2000, International journal of andrology.

[11]  W. Krause,et al.  Estimation of sperm morphology using a new GASA system , 2009, Andrologia.

[12]  D F Katz,et al.  Standardization and comparability of CASA instruments. , 1992, Journal of andrology.

[13]  P. Talbot,et al.  A triple-stain technique for evaluating normal acrosome reactions of human sperm. , 1981, The Journal of experimental zoology.

[14]  R. Menkveld,et al.  Acrosomal morphology as a novel criterion for male fertility diagnosis: relation with acrosin activity, morphology (strict criteria), and fertilization in vitro. , 1996, Fertility and sterility.

[15]  M. Béné,et al.  Lectin-binding sites on human sperm during acrosome reaction: modifications judged by electron microscopy/flow cytometry. , 1996, Archives of andrology.

[16]  W. Schill,et al.  Detection of human sperm acrosome reaction: comparison between methods using double staining, Pisum sativum agglutinin, concanavalin A and transmission electron microscopy. , 1997, Human reproduction.