Multiple Exposure Photography Method Research Articles

Citation classics in Fertility and Sterility, 1975–2004

Citation classics in Fertility and Sterility, 1975–2004

Superimposed Image Analysis System (SIAS) software: a new approach to sperm motility assessment

To apply Superimposed Image Analysis System (SIAS) software (Delta Sistemi, Rome, Italy) to the objective analysis of sperm motility. To test a new procedure for sperm motility assessment based on image superimposition. The system can superimpose six sequential frames onto a monitor producing a final image with a motion effect similar to the multiple exposure photography method, but bypassing the film processing stage. Viewing the final image of a complete series of six superimposed frames allows the evaluation of the percentage of motile spermatozoa and their kinetic characteristics. Compared with multiple exposure photography, SIAS has the following main advantages: [1] the procedure is quick; [2] projecting the first frame alone allows the easy evaluation of sperm concentration; and [3] the system allows quick and easy analysis and measurement of each sperm trajectory frame by frame, thus avoiding misleading interpretations because of crossing sperm trajectories, collisions, and overlapping of motile spermatoza with round cells or immotile spermatozoa. Superimposed image analysis is a new approach to sperm motility assessment. It seems to be useful particularly in selected situations, such as oligozoospermia or in cases of increased round cell concentration. In these situations, SIAS can even serve as a support or an alternative to the computer-aided sperm analysis system widely used for objective sperm analysis.

Rapid microcomputer-based analysis of semen characteristics from photographs taken by the MEP method.

Use of a microcomputer with the multiple exposure photography (MEP) method for rapid analysis of semen characteristics is described. The computer is programmed to calculate information fed from a digitizing tablet while scanning projected images of photographed sperm. In this way sperm concentration, percentage of motility, average velocity and frequency distribution of sperm velocity, as well as percentage of abnormal forms are immediately obtained. The method was found to be as accurate, less prone to error, and significantly less time consuming than the previous manual measuring method. It can greatly facilitate routine semen analysis in laboratories using the MEP method.

Objective evaluation of spermatozoa motility as influenced by fresh blood serum and normal saline

In order to investigate the effect of fresh blood serum and normal saline on the motility of ejaculated spermatozoa, semen samples were collected from 7 fertile males, aged 25–39 yr. Each sample was divided into 5 aliquots: one aliquot remained undiluted, and fresh blood serum from fertile men and normal saline were added to the others to produce 1:1 and 3: 1 dilutions. Spermatozoal motility was evaluated objectively, every hour and for 4 h after ejaculation, by the multiple exposure photography method. It was found that serum from fertile men as well as normal saline caused a significant increase of spermatozoal velocity. However, spermatozoal velocity under the influence of serum from fertile men changed with time in a different way than in the undiluted aliquot and those diluted with normal saline. Thus, the beneficial effect of fresh blood serum on spermatozoal velocity, cannot only be attributed to the lowering of seminal viscosity and the reduction of sperm concentration caused by dilution, but to various substances present in blood serum.

Effect of uremic serum on sperm motility: evaluation by multiple exposure photography.

The effect of serum from patients with severe chronic renal failure on the motility of ejaculated spermatozoa was objectively evaluated by the multiple exposure photography method. Uremic serum was not found to have a deleterious effect on the motility of ejaculated spermatozoa. On the contrary it caused a significant increase of spermatozoal velocity and a slighter increase in the percentage motility. These effects cannot be only attributed to the lowering of seminal viscosity and to the reduction of sperm concentration, but to various substances contained in blood serum.

Factors affecting sperm motility. VI. sperm viability under the influence of bacterial growth in human ejaculates

The influence of bacterial growth on human sperm motility and viability was evaluated objectively with the multiple-exposure photography method. Experimental semen specimens, obtained from normal donors bh nonaseptic means of masturbation, were incubated with antibiotics at room temperature or body temperature for 24 hours. Although bacteria, grew in control specimens, were totally eradicated in all antibiotic-treated specimens, no significant difference was found between these groups with regard to sperm motility throughout the time of incubation. Sperm survival was not inhibited, nor was it extended as a result of suppression of bacterial growth. In both groups, survival time was much shorter in specimens incubated at body temperature than in those kept at room temperature. Sperm motility was not affected after 2 hours of incubation of fresh specimens with concentrations of various pathogenic bacteria similar to those found in severe prostatitis. The question of whether the use of antibiotics in the treatment of asthenospermia per se has a prognostic value is discussed.

Distribution of normal and abnormal forms among motile, non-motile, live and dead human spermatozoa.

The distribution of normal and abnormal forms among motile, non-motile live, and dead spermatozoa was investigated . For this purpose samples from each specimen of 100 fertile and infertile patients was photographed twice: once with the aid of the multiple exposure photography (MEP) technique and again by an ordinary high-speed phase-contrast photomicrography. From enlarged projected photographed images, percentage of abnormal forms in both, non motile fraction and in the while sperm population were determined. By using a simple formula the percentage of abnormal forms among motile spermatozoa has been easily calculated. An average of 42% abnormal forms among all 100 analyzed specimens was found. However, this rate was not distributed equally among the two sperm populations: only 22% abnormal forms were among motile spermatozoa, contrary to 59% among non-motile spermatozoa. In 30 specimens that have been further analyzed by the combined supravital staining and the MEP method, percentage of abnormal forms among non-motile-live spermatozoa was found to be 32, and 67 among the fraction of dead spermatozoa. The importance of studying the distribution of normal forms among motile spermatozoa, and the question of any relationship of this parameter to the fertilization capacity of the semen, is discussed.

Studies on the components of kallikrein-kinin system and treatment of male infertility.

The kininase activity in the seminal plasma was identified with kininase II (angiotensin-converting enzyme) by thin layer chromatography and its activity of infertile patients was significantly lower than in normozoospermic patients. The amount of proteinase inhibitor in the seminal plasma was 7.0 ± 0.9 trypsin inhibitor unit/ml, however no correlation was found between its activity and semen quality.The effects of bradykinin and kallikrein on the velocity and motility of washed human spermatozoa were measured by multiple exposure photography method. The addition bradykinin at each concentration of 0.1ng/ml and 1.0ng/ml to the washed spermatozoa resulted in an increase of velocity and motility. The same effect as bradykinin on the spermatozoa was found in the presence of 0.1 KU/ml and 1.0 KU/ml of kallikrein.In order to determine the effect of kallikrein on the concentrations of seminal fluid components, infertile men were treated orally for 6 months. Following determinations of volume, motility and sperm density, the concen-trations of kininase II, proteinase inhibitor, lactate dehydrogenase and acid phosphatase were measured. Sperm motility and density were increased dur-ing treatment. Mean values of the parameters in the ejaculate unchanged, with the exception of kininase II, which exhibited significantly higher con-centration under treatment.Of a total of 131 men treated, 23 pregnancies (17.6%) is established be-tween the first and the sixth month.Thus, present work shows that the components of kallikrein-kinin system participate in spermatogenesis and sperm motility.

Factors Affecting Sperm Motility. II. Human Sperm Velocity and Percentage of Motility as Influenced by Semen Dilution

Spermatozoal velocity and percentage of motility were analyzed objectively with the multiple exposure photography method before and after specimens from fertile and infertile men were diluted in their own seminal plasma or normal saline. No significant change in percentage of motility was found in samples diluted up to 1:6 in both kinds of diluents. However, a significant relative increase (up to 25% of the original velocity) was found when a specimen was diluted with its own seminal plasma, and an even greater increase (up to 37% of the original velocity) was found when it was diluted with saline. Compared with undiluted specimens, there was no delayed effect on spermatozoal motility when semen was diluted with saline after up to 4 hours' incubation time. Contrary to the findings in animal and human semen described by others, there was no deleterious effect on sperm motility with this kind and rate of dilution and duration of time. The assumption that the increase in sperm velocity caused by dilution is not excitatory but is due only to a decrease of seminal fluid viscosity and a reduced number of spermatozoa which interfere with sperm free movement is discussed. We recommend evaluation of spermatozoal motility in diluted specimens in addition to evaluation of the original specimen in any routine semen analysis in order to determine true spermatozoal motility potential under optimal conditions.

Sperm Velocity and Percentage of Motility in 100 Normospermic Specimens Analyzed by the Multiple Exposure Photography (MEP) Method

Specimens from 100 fertile or potentially fertile men with sperm counts above 20 million/ml were analyzed objectively by the new multiple exposure photography method (MEP). The following results (means and standard deviations) were obtained: sperm concentration, 66 million/ml +/- 47; percentage of motile spermatozoa, 45% +/- 18; speed of motile spermatozoa, 30.3 micrometer/second +/- 5.5; abnormal forms, 29% +/- 10.2; average motility index (product of sperm speed and fraction of motile spermatozoa), 14 +/- 7.5. Low correlations were found between sperm concentration, percentage of motility, and spermatozoal velocity. Moderate correlations were found between percentage of motility and spermatozoal velocity. Negative moderate correlations were found between abnormal forms and spermatozoal velocity as well as between abnormal forms and percentage of motility. The differences between some of our results and those obtained by others are discussed.