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Takumi Tsuji

Bio: Takumi Tsuji is an academic researcher from Kobe University. The author has contributed to research in topics: Sperm. The author has an hindex of 1, co-authored 1 publications receiving 11 citations.
Topics: Sperm

Papers
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Journal ArticleDOI
TL;DR: Semen samples from 104 men attending a male infertility clinic were used to evaluate the reproducibility of results and the carryover rate with the S-FCM by performing between- and within-run imprecision analyses, showing that automated analyses were highly reproducible and the Carryover rate was 0.17% or less.
Abstract: The S-FCM uses flow cytometry technology to measure sperm concentrations. Semen samples from 104 men attending a male infertility clinic were used to evaluate the reproducibility of results and the carryover rate with the S-FCM by performing between- and within-run imprecision analyses. In addition, sperm concentrations measured with the S-FCM were compared with those obtained by manual analyses with the Makler chamber and the improved Neubauer hemacytometer. The results showed that automated analyses with the S-FCM were highly reproducible and the carryover rate was 0.17% or less. In within-run imprecision assays, the coefficients of variation for the S-FCM were less than 5% at all sperm concentrations, while those for the Makler chamber were between 17.7% and 28.7% at lower sperm concentrations. The overall correlation between values measured with the S-FCM and those measured with the Makler chamber and improved Neubauer hemacytometer was excellent, but at lower sperm concentrations the correlation was lower. The S-FCM performed sperm concentration analyses in 110 seconds compared with 5 minutes for the Makler chamber and 10 minutes for the improved Neubauer hemacytometer. The S-FCM is suitable for quantitative measurement of lower sperm concentrations. Sperm counts are an essential step in the evaluation of male fertility and usually are performed by microscopic examination of semen by trained personnel. The World Health Organization (WHO) recommends the use of a hemacytometer for determining sperm concentrations in semen, 1 and several methods of using these devices have been described. 1-3 However, the accuracy and reproducibility of analytic data obtained with hemacytometers reportedly are not good enough to compare results from different institutions. 4-6 A number of studies have shown that manual counting is associated with the greatest potential for gross analytic errors in measuring sperm concentrations. 4-8 Consequently, computer-assisted semen analyzers were developed to overcome the subjectivity of sperm counting, but most of these analyzers use some sort of counting chamber or hemacytometer that is itself a source of intrinsic errors. When using a counting chamber or hemacytometer, only spermatozoa located in it can be counted, and only very small quantities of specimens are used to estimate the sperm concentration in 1 mL of semen. Recently we developed a new device (S-FCM) for measuring sperm concentrations that uses flow cytometry technology and an argon laser as the light source. To improve accuracy, the S-FCM device uses 90 times greater quantities of semen than are used with a counting chamber or hemacytometer. In this study, we compared the accuracy and reproducibility of the S-FCM with that of the Makler sperm counting chamber. We also evaluated the feasibility of using the S-FCM in clinical settings by comparing the analytic data with those obtained by using the improved Neubauer hemacytometer or the Makler chamber.

12 citations


Cited by
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Journal ArticleDOI
TL;DR: It is concluded that duplicate counts by at least two technicians is recommended to achieve high precision but, that particular caution should be exerted with regard to the precision and accuracy of the used counting device.

94 citations

Journal ArticleDOI
TL;DR: A new flow cytometric method has been developed to rapidly determine sperm concentration and viability in semen from bulls and boars, and it was found that this system was two and four times more accurate than the spectrophotometer and the electronic cell counter, respectively.
Abstract: A new flow cytometric method has been developed to rapidly determine sperm concentration and viability in semen from bulls and boars. Sperm viability was determined on the basis of staining with SYBR-14 and propidium iodide (PI), and this allowed detection of live (membrane-intact) sperm, dying (moribund) sperm, as well as dead cells. Fluorescent microspheres (beads) were used to determine sperm concentration. The use of SYBR-14 at 50 nM and PI at 12 μM in combination with the FACSCount diluent in the counting tubes resulted in a uniform staining after 2.5–5 minutes at room temperature. Reagent staining was reproducible enough to allow subsequent semiautomated analysis of data using Attractors software. In experiment 1, this method was used to analyze semen from boars, rams, rats, rabbits, humans, and turkeys. In experiment 2, Attractors analysis was performed by the FACSCount AF flow cytometer, and sperm concentration determination with this system was compared with results obtained by a spectrophotometer and an electronic cell counter, which is routinely used by bull artificial insemination centers. When compared to microscopic counting in a hemocytometer, the FACSCount AF flow cytometer was two and four times more accurate than the spectrophotometer and the electronic cell counter, respectively. In addition, the FACSCount AF flow cytometer determined both sperm concentration (coefficient of variation 3.3%) and sperm viability (coefficient of variation 0.7%) with high precision.

74 citations

Journal ArticleDOI
TL;DR: The automated SQA-V analyzer is more precise and shows the ability to accurately classify normal versus abnormal sperm morphology and can be used interchangeably with manual semen analysis methods for examining sperm concentration and motility.

46 citations

Journal ArticleDOI
TL;DR: The information covered in the article includes sample preparation and the use of manual counts, spectrophotometers, computer-assisted semen analysis, NucleoCounter, and flow cytometry.

34 citations

Journal ArticleDOI
TL;DR: Flow cytometry can result in an overestimation of both bead counting and sperm concentration, suggesting that flow cytometry is an inappropriate method for sperm counting, especially in the case of azoospermia.
Abstract: It has been reported that flow cytometry can be used as a reference procedure to determine sperm concentrations in quality control schemes in andrology laboratories, but there are no convincing quality control data. To understand comprehensively whether flow cytometry can be used to determine sperm concentration, sperm concentrations of 85 human semen samples were detected using three different methods, namely flow cytometry, computer-assisted semen analysis (CASA) and manual counting with a cell-VU chamber. The bead concentrations of both low [(18+/-2.5)x10(6)/mL] and high [(35+/-5)x10(6)/mL] pre-calibrated standard latex bead solutions were also determined with flow cytometry. The results showed that bead concentrations of both low and high pre-calibrated standard latex bead solutions counted five times with flow cytometry were (21.37+/-0.85)x10(6)/mL and (45.95+/-1.76)x10(6)/mL, respectively. Coefficient variances (CVs) and relative errors (REs) were 4%, 15.51% and 3.84%, 31.3% for low and high latex bead solutions, respectively. The overall correlation between values measured with flow cytometry and values measured with the cell-VU chamber and the CASA system was significant. However, flow cytometry overestimated the sperm concentration by 109% compared to the results with the cell-VU chamber. Moreover, for the azoospermic samples analysed, the sperm concentration was estimated at 0.12 (range from 0.04 to 0.24)x10(6)/mL. In conclusion, the data demonstrated that flow cytometry can result in an overestimation of both bead counting and sperm concentration, suggesting that flow cytometry is an inappropriate method for sperm counting, especially in the case of azoospermia.

18 citations