[Influence of different filling method-related sperm counting chambers and structural factors of disposable chambers on sperm motility].

Objective: To evaluate the effects of different filling method-related sperm counting chambers and the structural factors of Leja counting chambers on sperm motility using computer-assisted sperm analysis (CASA). METHODS: Using drop-filled Makler, capillary-loaded Leja and structurally modified Leja sperm counting chambers, we measured sperm concentration, the percentages of progressively motile sperm (PMS) and non-progressively motile sperm (NPMS), total sperm motility, curvilinear velocity (VCL), average path velocity (VAP), straight line velocity (VSL), beat-cross frequency (BCF), linearity (LIN), wobble (WOB) and straightness (STR) in the semen samples of 76 males by CASA and compared them between different chambers. RESULTS: The drop-filled Makler sperm counting chamber achieved remarkably higher PMS, NPMS, total sperm motility, VCL and VAP than the Leja chambers (P < 0.05). There were no statistically significant differences in VSL, BCF, LIN, WOB and STR between the Makler and Leja chambers (P > 0.05), or in sperm concentration, PMS, NPMS and total sperm motility between the capillary-loaded and structurally modified Leja counting chambers (P > 0.05). The ground edge and thickness of the coverslip of the Leja counting chamber produced no significant inference on the kinetic sperm parameters (P > 0.05). CONCLUSIONS: The drop-filled sperm counting chamber achieves significantly higher sperm motility and kinetic parameters than the capillary-loaded Leja chamber. The structural factors such as the ground edge and thickness of the coverslip of the Leja counting chamber do not influence the analysis of sperm parameters.

Accuracy Evaluation of The Depth of Six Kinds of Sperm Counting Chambers for both Manual and Computer-Aided Semen Analyses.

BACKGROUND: Although the depth of the counting chamber is an important factor influencing sperm counting, no research has yet been reported on the measurement and comparison of the depth of the chamber. We measured the exact depths of six kinds of sperm counting chambers and evaluated their accuracy. MATERIALS AND METHODS: In this prospective study, the depths of six kinds of sperm counting chambers for both manual and computer-aided semen analyses, including Makler (n=24), Macro (n=32), Geoffrey (n=34), GoldCyto (n=20), Leja (n=20) and Cell-VU (n=20), were measured with the Filmetrics F20 Spectral Reflectance Thin-Film Measurement System, then the mean depth, the range and the coefficient of variation (CV) of each chamber, and the mean depth, relative deviation and acceptability of each kind of chamber were calculated by the closeness to the nominal value. Among the 24 Makler chambers, 5 were new and 19 were used, and the other five kinds were all new chambers. RESULTS: The depths (mean ± SD, µm) of Makler (new), Macro and Geoffrey chambers were 11.07 ± 0.41, 10.19 ± 0.48 and 10.00 ± 0.28, respectively, while those of GoldCyto, Leja and Cell-VU chambers were 23.76 ± 2.15, 20.49 ± 0.22 and 24.22 ± 2.58, respectively. The acceptability of Geoffrey chambers was the highest (94.12%), followed by Macro (65.63%), Leja (35%) and Makler (20%), while that of the other two kinds and the used Makler chamber was zero. CONCLUSION: There existed some difference between the actual depth and the corresponding nominal value for sperm counting chambers, and the overall acceptability was very low. Moreover, the abrasion caused by the long use, as of Makler chamber, for example, may result in unacceptability of the chamber. In order to ensure the accuracy and repeatability of sperm concentration results, the depth of the sperm counting chamber must be checked regularly.

Comparison of the semen analysis results obtained from two branded computer-aided sperm analysis systems.

The study evaluated the comparability of two branded computer-aided sperm analysis (CASA) systems commonly used in andrology laboratories in China. The same semen sample was analysed using two branded CASA systems (WLJY-9000 and CFT-9200) by one well-trained technician. Results of semen analysis obtained from two branded CASA systems were then compared. The accuracy of counting results of CASA systems was evaluated using latex bead solutions with known concentrations of (35 ± 5) × 106 ml⁻¹ and (18 ± 2.5) × 106 ml⁻¹. There were significant differences in all parameters (P < 0.01) except for LIN and WOB. The counting results of CFT-9200 were close to the standard solutions [(38.86 ± 3.79) × 106 ml⁻¹ and (19.03 ± 1.99) × 106 ml⁻¹], while those of WLJY-9000 were underestimated [(28.53 ± 2.06) × 106 ml⁻¹ and (14.62 ± 0.95) × 106 ml⁻¹]. But the coefficient of variation of WLJY-9000 was lower than that of CFT-9200 (7.22%, 6.50% vs. 9.82%, 10.46%). It is concluded that factors such as parameter settings and evaluation algorithms could significantly affect the results obtained from these two branded CASA systems. Great attention should also be paid to the quality control in semen analysis with CASA.

Effect of Ethane 1,2-Dimethane Sulfonate(EDS) on the Testicular Expression of Steroidogenesis-related Genes and Epididymal Sperm Count in the Adult Rat / 대한남성과학회지

PURPOSE: Ethane 1,2-Dimethane sulfonate (EDS), an alkylating agent, has been widely used to create the testosterone withdrawal rat model. The present study was carried out to test the effect of EDS administration on the expression of steroidogenesis-related genes in the rat testis and on epididymal sperm counts. MATERIALS AND METHODS: Adult male Sprague-Dawley rats (300~350 g B.W.) were injected with a single dose of EDS (75 mg/kg, i.p.) and sacrificed on days 0, 7, 14, 21, 28, 35, 42, and 49. Tissue weights (testis, epididymis and seminal vesicle) were measured, and serum LH levels were determined by specific radioimmunoassay. The transcriptional activities of LH receptor (LH-R), 3beta-hydroxysteroid dehydrogenase (3beta-HSD), and steroidogenic acute regulatory protein (StAR) were evaluated by semi-quantitative RT-PCR. RESULTS: Weights of the reproductive and accessory organs declined progressively after the EDS treatment (weeks 1~3). After this, the decrease stopped, with a gradual return towards normal. Full recovery was observed in testis and seminal vesicle evaluations on weeks 5 and 6, respectively. Only 70% recovery was found in epididymis during weeks 5~7. A more dramatic drop was observed in caput epididymal sperm count, and the maximum recovery was 40% on week 7. Serum LH level increased significantly on week 2 after EDS treatment, then gradually decreased during weeks 3~5. The transcripts for the steroidogenesis-related genes in testis declined sharply during weeks 1~2, then returned to normal on week 4. CONCLUSIONS: Our results demonstrate that EDS might directly induce severe damage, such as tissue destruction and decreased sperm counts, in epididymis compared to those in testis and seminal vesicles. Changes in the activities of testicular steroidogenesis-related genes caused by abrupt death and repopulation of Leydig cells in EDS-treated rats were in good correlation with other parameters shown in this and previouslypublished data. Taken together, the EDS injection model might be useful to understand not only the mechanism of differentiation of testicular somatic and germ cells but also the function of the epididymis in the aging process.