World J Urol (1986) 4:65-70 Woad a o , ~ of 7 Urology © Springer-Verlag 1986 Spermiometrics: the method and its clinical application* K. Scheiber 1, A. Schmassmann 2, H. P. Rohr 2, P. J. Oefner 1, M. Gotwald 1 and G. Bartsch 1 1 Department of Urology, Anichstrasse 35, A-6020 Innsbruck, Austria 2 Department of Pathology, CH-4031 Basel, Switzerland Summary. Objective and reproducible determinations of sperm motility and morphology are achieved using a semi-automatic image analysis system under optomanu- al control. The method was used to evaluate the short- and long-term effects of the operative treatment of vari- cocele on sperm density, motility and velocity in 27 men with left-sided varicosities. Whereas a significant im- provement Co = 0.0001) in sperm density was found within 3 months after surgery, almost no increases in sperm motility and velocity occurred. The actual post- operative improvement in sperm density did not cor- relate with the preoperative sperm counts (r-- 0.291). Between the preoperative values of motility and velocity and the postoperative improvement, however, a negative correlation of-0.431 and -0.759, respectively, was found. Considering the 21 patients, who were followed up for a minimum of 9 months, a sustained increase in sperm density, motility and velocity of at least 50% was observed in 14 (66.70/0), 7 (33.3O/o) and 5 (23.80/0), respec- tively. In general, the better results were obtained in the group with the poorer preoperative semen character- istics. However, only 4 patients (190/0) showed a lasting improvement in all 3 parameters. Even more interesting was the sustained decrease in sperm velocity observed in 9 men (42.9 %), who had rather good preoperative values. It is concluded, that a timely operative treatment ofvari- cocele might prevent further impairment of semen quality. However, more research on the natural course of varicocele is necessary. Method Introduction Sperm density, motility and morphology are among the most important clinical parameters for estimating the fertilizing capacity of semen. However, present semen analysis is limited by subjective evaluation and insuf- ficient information. Accordingly, the indication for appropriate treatment is difficult to ascertain, and the * Supported by the Fonds zur F6rderung der wissenschafllichen Forschung No. P3278, Austria success of treatment can hardly be appraised. Subjective evaluation makes standardization and comparison be- tween results of different laboratories difficult. Recently, low-cost semi-automatic image analysis systems have been developed, allowing quick and pre- cise measurement of sperm motility and morphology as well as statistical analysis of measured data [14]. Technique Technical set-up. The determination of sperm motility and morphology is achieved by means of the same tech- nical set-up (Fig. 1), which consists of a microscope with a drawing-tube (Diavar, C. Reichert AG, Wien, Austria), a digitizer tablet (Kontron) and a cursor interfaced to the semi-automatic image analysis system MOP AM 02 (Kontron, Munich, FRG). A light-emitting diode, which is mounted on the cursor and visible as a bright red pilot light in the microscopic field, permits the convenient tracing of spermatozoa on the digitizer tablet under opto- manual control. The statistical calculations are per- formed by the microcomputer HP 9815 A (Hewlett- Packard, Loveland, CO 80537, USA). For motility measurements, phase contrast or dark- field illumination is recommended. Morphology meas- urements are performed under bright-field illumination using a planapochromatic objective (100:1) and an addi- tional lens, which allows a final magnification on the digitizer tablet of at least 5000. Motility. Motility measurements are performed approxi- mately 1 hour after semen collection at an ambient tem- perature. If the sperm density exceeds 100 mill/ml the semen has to be diluted with Normon-Johnson-Solution I, which minimally influences motility [7]. For the first experiments, a microchamber, which is exactly 1 ~tm deep, was used. Since the handling and cleaning of the device did not permit the analysis of more than one semen sample under routine conditions, it was abandon- ed in favor of an ordinary glass slide, one drop of semen being covered with a coverglass. As outlined in the sec- tion on the clinical application of spermiometrics, this adjustment did not influence the results significantly.