COMPUTER AUTOMATED MORPHOMETRIC ANALYSIS OF BULL SPERM HEADS

Morphological type classification of spermatozoa is an important compo nent of the modern semen evaluation; however, current methods of analy sis are subjective and highly variable between technicians. To reduce the subjectivity and thus variability of sperm morphology assessment, computer automated sperm head morphology analysis (ASMA) has been deve loped. Previous studies have shown the importance of standardizing ASM A procedures to optimize accuracy. The objective of this study was to standardize ASMA procedures for evaluating bull sperm heads. Semen fro m 10 fertile bulls was used to standardize procedures for optimal anal ysis of bull spermatozoa. Sample preparation methods, sperm staining m ethods and microscopic magnifications were compared. Semen samples tha t were diluted to a standard concentration of 200x10(6) sperm/ml were more efficiently analyzed than raw samples. A modified GZIN staining p rocedure, incorporating rose bengal as an acrosomal stain, was used fo r accurate ASMA at a magnification of x 60. The mean morphometric meas urements for ail bulls were the area (27.30 mu M), perimeter(25.36 mu M), length(8.65 mu M), width(4.40 mu M) and width/length (0.50). Withi n the analyses, coefficients of variation ranged from 3.45% for length to 8.52% for area. The ASMA system correctly digitized sperm heads 97 % of the time. Results of this study indicate that bull sperm heads ca n be accurately analyzed using current standard procedures of ASMA tec hnology.