Validation of the FACSCount AF system for determination of sperm concentration in boar semen

Hansen, C., Christensen, P., Stryhn, H., Hedeboe, A. M., Rode, M. and Boe-Hansen, G. (2002) Validation of the FACSCount AF system for determination of sperm concentration in boar semen. Reproduction in Domestic Animals, 37 6: 330-334. doi:10.1046/j.1439-0531.2002.00367.x


Author Hansen, C.
Christensen, P.
Stryhn, H.
Hedeboe, A. M.
Rode, M.
Boe-Hansen, G.
Title Validation of the FACSCount AF system for determination of sperm concentration in boar semen
Journal name Reproduction in Domestic Animals   Check publisher's open access policy
ISSN 0936-6768
1439-0531
Publication date 2002-12
Sub-type Article (original research)
DOI 10.1046/j.1439-0531.2002.00367.x
Open Access Status Not yet assessed
Volume 37
Issue 6
Start page 330
End page 334
Total pages 5
Place of publication Berlin, Germany
Publisher Wiley-Blackwell Verlag GmbH
Language eng
Formatted abstract
A flow cytometric method has been developed for rapid determination of sperm concentration in semen from various mammalian species.* All cells containing DNA are stained with SYBR-14 or propidium iodide (PI) and sperm concentration is determined in relation to an internal standard of fluorescent microspheres (beads). Satisfactory staining can be achieved within 2-3 min and the following flow cytometric analysis on the FACSCount AF System rapidly provides the user with a precise and accurate assessment of the sperm concentration. In this study, the FACSCount AF System and Sperm Counting Reagent (BD Biosciences) was compared with microscopic counting using a Bürker-Türk haemocytometer. In addition, sperm concentration was determined using the Corning 254 spectrophotometer which is used routinely by Danish artificial insemination stations for boars. The results show that the agreement between flow cytometry and microscopic counting is very high. The slope for the regression line was 1.12 (SE = 0.03) with an estimated intercept with the Y-axis of 22 × 106 sperm/ml (SE = 10 × 106 sperm/ml) and an estimated error of the model of 10 × 106 sperm/ml. For the spectrophotometer, the slope of the regression line was 1.09 (SE = 0.07) with an estimated intercept of 137 × 106 sperm/ml (SE = 25 × 106 sperm/ml). The average error made by the spectrophotometer was 55 × 106 sperm/ml. In addition, the results obtained using flow cytometry was highly repeatable (CV = 2.7%) in comparison with the spectrophotometric method (CV = 6.3%). These results indicate that the FACSCount AF System is a valuable tool for precise and accurate assessment of sperm concentration in boar semen and that use of this system may lead to production of more uniform insemination doses containing a specific number of sperm per dose.
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: School of Veterinary Science Publications
 
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