Feasibility study for the centralized measurement of sperm concentration
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Despite several efforts to standardize methods of semen analysis, sperm count is known to be subject to large interlaboratory differences. This is especially a problem in multicentre clinical trial settings and protocols for the preparation of semen for centralized assessment of sperm concentration are suggested here. The stability of semen has been tested after fixation with formalin at different dilutions and at different temperatures for different sperm concentrations. Sperm concentrations in formalin-fixed semen (at dilutions 1 + 0.5 to 1 + 19) were stable (<20% difference from original) at all concentrations (0.1 x 10(6)/mL to 100 x 10(6)/mL) for at least 5 days at room temperature and 4 degrees C. Prolonged stability up to 5 weeks at 4 degrees C was demonstrated for the lower dilutions (1 + 0.5 and 1 + 1). Freezing of fixed semen samples was unacceptable. These results illustrate that centralized assessment of sperm concentrations is feasible.ABSTRACTObjective: This study aimed to determine the relationship between the addition of Glutathione to sperm freezing medium and cryosurvival rate of sperm motility and viability in asthenozoospermic patients.Methodology: fifty nine semen samples of asthenozoospermic patients were obtained from the male partner of infertile couples attending the fertility center of al-sadder medical teaching city in Al-Najaf during the period 1st October 2016 until 1st March 2017.Each semen sample was divided into four equal parts, volume of each part is 0.5 ml: first part (before activation), second part (after activation), third part (cryopreservation with sperm freezing medium for one month), and fourth part (cryopreservation with sperm freezing medium plus glutathione for one month ) .Sperm concentration , sperm progressive motility, sperm viability and sperm normal morphology were counted in first two parts, cryosurvival rate of sperm motility, viability was counted in third and fourth parts .Statistical analysis of the study performed by using Independent t-test and paired test to assess the differences between two groups .Results: After sperm activation by using the swim up technique ,the study results showed significant increase (P < 0.01) of progressive sperm motility, normal sperm morphology and sperm viability, while there were a significant decrease (P < 0.01) of sperm concentration. cryosurvival rate of sperm motility and viability were significant decrease (p < 0.01) after cryopreservation and thawing processes while the results were revealed significant improvement in cryosurvival rate of sperm motility, and viability ( p < 0.01 ) after the addition of glutathione to the sperm freezing medium.Conclusion: it can conclude from this study that addition of glutathione to sperm freeze medium improves. (p < 0.01) cryosurvival rate of sperm motility and viability during cryopreservation and thawing processes . Recommendations: the study recommended to addition 50 microliter of glutathione to sperm freezing media in concentration 1 mM .
Semen Analysis
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The thesis introduce a solution of using DataGrid coned and ComboBox control to realize DataBase of querying
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The removal of rival sperm from a female's sperm storage organ acts as a strong sperm competition avoidance mechanism, which has been reported only in internally fertilizing species and not at all in externally fertilizing species. This study demonstrated for the first time that nest-holding males of Bathygobius fuscus , an externally fertilizing marine fish, remove the sperm of rival sneaker males from the spawning nest by exhibiting tail-fanning behaviour within the nest. Males showed tail-fanning behaviour when semen was artificially injected into the nest but not when seawater was injected, and in open nests this behaviour resulted in higher paternity rates for the focal male. The sperm removal behaviour entails the risk of removing their own sperm; therefore, additional sperm release behaviour is likely necessary to benefit from the sperm removal effect. Consistent with this, males increased post-fanning sperm release behaviour more in the semen than in the seawater injection treatment. Moreover, males who had removed sperm for a longer time spent more time releasing sperm after the removal, suggesting that the additional sperm release behaviour compensated for the loss of their own sperm. These results suggest that sperm removal behaviour is not restricted to internally fertilizing organisms and deserves further investigation in this and other species.
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Evaluation of rat sperm by flow cytometry: Simultaneous analysis of sperm count and sperm viability.
In this study, we conducted a simultaneous analysis of sperm count and viability in rats by flow cytometry (FCM). Epididymal fluids were taken from the caudal epididymis of 12 to 13 week-old Sparague-Dawley rats. The fluids were weighed and mixed with Dulbecco's phosphate buffered saline (D-PBS). Propidium iodide, which can stain only dead sperm, was used to distinguish viable and dead sperm. The sperm count and viability analyzed by FCM were 1.28×106/mg and 78.0%, respectively. These values were consistent with the corresponding values (1.39×106/mg and 81.0%) that were directly determined microscopically in the fluids of the same sample. In addition, when the original mixture containing sperm was diluted two times and four times with D-PBS, or was diluted two times with D-PBS containing only killed sperm, the sperm count and viability determined by FCM also correlated well with the sperm count (r=0.96, P<0.01) and sperm motility (r=0.99, P<0.01) by direct microscopic observation, respectively. In conclusion, the present flow cytometric analysis would be practical for the simultaneous determination of sperm count and viability in rat epididymal fluids.
Propidium iodide
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Six-week old SD-Slc male rats were treated for 4 weeks with compounds known to induce toxicological changes in male reproductive organs (pyridoxine in saline, 500 mg/kg/day, i.p.) or sperm (trimethylphosphate in distilled water, 100 mg/kg/day, p.o.). Each sperm sample taken from the cauda epididymis was analyzed with flow cytometry for the evaluation of sperm viability and counts. Sperm motility and morphology by microscopical observation, and histopathological examination of reproductive organs were also performed for estimating the adverse effects of each compound on spermatogenesis and sperm. While a decrease in sperm motility was noted for the trimethylphosphate group, the low motile sperm was evaluated as being viable sperm, not moribund, with flow cytometry. In the pyridoxine group, microscopical observation revealed morphological changes of sperm and a decrease of motility. The present sperm analysis with flow cytometry also suggested morphological changes reflected by dot plot as well as decrease of sperm viability and counts. These results indicated that this procedure led to profound findings in the compound-treated animals, with evaluation as viable sperm, not moribund, in a low motile sperm sample, and suggesting morphological changes in the dot plot of flow cytometry.
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Semen Analysis
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This paper analyzed the DataGrid Web Server Control. DataGrid is one of the most popular control of ASP .NET which is used to render data to a Web page in tabular form. This paper provides two types of typical usage of DataGrid.
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Viable, healthy sperm are preferred for oocyte fertilization with intracytoplasmic sperm injection. Currently, motility is the most widely applied measure of sperm viability. However, with this criterion, viable but immotile sperm are overlooked as candidates for micromanipulation. Supravital stains identify viable sperm but may be toxic to the gamete or embryo. We tested the hypothesis that hypo-osmotic swelling, developed to assess sperm membrane integrity, can accurately determine sperm viability. Thawed sperm from 12 fertile donors were exposed to a hypo-osmotic solution and to two supravital stains. A total of 2,010 sperm were assessed for tail coiling after a 30-min exposure to hypoosmotic solution. By supravital stains, 31% of thawed sperm were viable; 23% showed tail coiling. Among coiled-tail sperm, 100% were viable by supravital stain. As a measure of viability, tail coiling exhibited a sensitivity of 30%, specificity of 100%, and positive predictive value of 100% compared to supravital stains. With a 60-min hypo-osmotic incubation, the specificity (89%) and positive predictive value (78%) decreased significantly (P < 0.05). Therefore, hypo-osmotic swelling accurately detects viable sperm among a nonmotile population. Assay accuracy, however, is very sensitive to the incubation time in hypo-osmotic solution.
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Dichlorofluorescein
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