Automated Sperm Morphometry Analysis (ASMA) in the Rabbit

Abstract

ABSTRACT: New methods of specimen preparation were developed and a new method of objective, automated sperm morphometry analysis (ASMA) was performed to reduce the technical variation in the rabbit sperm morphology assay. The optimal staining procedure was a modified GZIN stain, which allowed the ASMA instrument to accurately recognize the distal end of the sperm head and to achieve the highest sperm recognition rate (94%). Washing and resuspending sperm to a standard concentration increased the number of sperm per microscopic field in low‐concentration samples and reduced the field‐to‐field variation in all samples. Washing also decreased the number of sperm recognition errors by the ASMA instrument. Mean metric measurements for all sperm were: length, 7.38 μm; width, 3.91 μm; width/length, 0.53; area, 22.10 μm; and perimeter, 19.20 μm. Within‐specimen coefficients of variation (CVs) ranged from 0.8% to 5.5% and between‐animal CVs ranged from 2.7% to 7.5%. The use of standard specimen preparation techniques and ASMA technology can significantly reduce the technical variation in the rabbit sperm morphology assay. 1995 American Society of Andrology