RAPID, OBJECTIVE AUTOMATED SYSTEM TO ANALYZE RABBIT SEMEN

THE RABBIT IS A VALUABLE MODEL FOR REPRODUCTIVE TOXICITY STUDIES AND IS ESPECIALLY USEFUL FOR ACUTE, CHRONIC, AND REVERSAL STUDIES IN THE MALE. ITS SEMEN IS DIFFICULT TO ANALYZE IN COMPUTER SYSTEMS DUE TO THE PRESENCE OF PARTICLESEASILY MISTAKEN FOR NONMOTILE SPERM. THIS PROJECT WILL DEVELOP A PROTOTYPE AUTOMATED ANALYZER FOR OBJECTIVELY EVALUATING RABBIT (AND OTHER) SEMEN, IN WHICH THE ERRORS CAUSED BY INTERFERING GRANULES AND BY HIGH SPERM VELOCITY ARE ELIMINATED. THIS PROTOTYPE WILL USE A STROBOSCOPIC PHASE-CONTRAST OPTICAL SYSTEM SPECIALLY ADAPTED BOTH TO DISCRIMINATE BETWEEN SPERM AND GRANULES AND TO REDUCE VELOCITY-INDUCED IMAGE BLURRING TO INSIGNIFICANT LEVELS. MORPHOMETRIC ANALYSIS WILL ALSO BE USED AS AN ADDITIONAL DISCRIMINATOR IFNECESSARY. THE CRITICAL EXPERIMENTS TO VALIDATE THE ALGORITHMS FOR GRANULE DISCRIMINATION WILL BE PERFORMED BY HARVESTING GRANULES AND ADDING THEM BACK, IN KNOWN AMOUNTS, TO GRANULE-FREE, POOLED SEMEN SAMPLES. WITH THESE IMPROVEMENTS, IT SHOULD BE POSSIBLE TO ACCURATELY, CONVENIENTLY, AND RAPIDLY DETERMINE SPERM CONCENTRATION, MOTILITY, AND VELOCITY.