Journal of Andrology
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow An erratum has been published
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Christensen, P.
Right arrow Articles by Godfrey, W. L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Christensen, P.
Right arrow Articles by Godfrey, W. L.
Journal of Andrology, Vol. 25, No. 2, March/April 2004
Copyright © American Society of Andrology

A Flow Cytometric Method for Rapid Determination of Sperm Concentration and Viability in Mammalian and Avian Semen

PREBEN CHRISTENSEN*, JENS P. STENVANG{dagger} AND WILLIAM L. GODFREY{ddagger}

From the * Department of Clinical Studies, Section for Reproduction, Royal Veterinary and Agricultural University, Frederiksberg, Denmark; {dagger} Bartholin Instituttet, Copenhagen, Denmark; and {ddagger} BD Biosciences Immunocytometry Systems, San Jose, California.

Correspondence to: Dr Preben Christensen, Department of Clinical Studies, Section for Reproduction, Royal Veterinary and Agricultural University, Dyrlaegevej 68, 1870 Frederiksberg C, Denmark (e-mail: PC{at}kvl.dk).


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.

     Key words: Cytometry, spermatozoa, count, vitality, precision, accuracy




This article has been cited by other articles:


Home page
 Hum ReprodHome page
S. Perticarari, G. Ricci, M. Granzotto, R. Boscolo, C. Pozzobon, S. Guarnieri, A. Sartore, and G. Presani
A new multiparameter flow cytometric method for human semen analysis
Hum. Reprod., February 1, 2007; 22(2): 485 - 494.
[Abstract] [Full Text] [PDF]

Home page
 J AndrolHome page
P. Christensen, D. Boelling, K. M. Pedersen, I. R. Korsgaard, and J. Jensen
Relationship Between Sperm Viability as Determined by Flow Cytometry and Nonreturn Rate of Dairy Bulls
J Androl, January 1, 2005; 26(1): 98 - 106.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 2004 by The American Society of Andrology.