a1 Yves Ménézo. Laboratoire Clément, 17 avenue d’Eylau, 75016 Paris and 8 avenue Henri Barbusse, 93150 Le Blanc Mesnil, France
a2 Laboratoire Clément, 17 avenue d’Eylau, 75016 Paris and 8 avenue Henri Barbusse, 93150 Le Blanc Mesnil, France.
a3 Urology Department, American Hopsital of Paris, 92200 Neuilly-sur-Seine, France.
a4 Beckman Coulter France, Villepinte, 95942 Roissy, France.
a5 Biological Reproductive Products (BRP), Lugano, Switzerland.
The impact of sperm DNA fragmentation on assisted reproductive technology (ART) successes, in terms of outcome, is now established. High levels of DNA strand breaks severely affect the probability of pregnancy. The importance of sperm nucleus condensation in early embryogenesis and, subsequently, on the quality of the conceptus is now emerging. In this article we have compared in situ analyses with terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labelling (TUNEL) (for DNA fragmentation) with aniline blue (AB) (for nucleus decondensation), versus flow cytometry (FC) after acridine orange staining, in a double-blinded analysis. In our hands, TUNEL and acridine orange give perfectly comparable results. For decondensation the results are also comparable, but the double-stranded green fluorescence obtained with acridine orange seems to slightly underestimate the decondensation status obtained with AB.
(Received December 22 2013)
(Revised March 31 2014)
(Accepted April 01 2014)
(Online publication July 03 2014)
c1 All correspondence to: Yves Ménézo. Laboratoire Clément, 17 avenue d’Eylau, 75016 Paris and 8 avenue Henri Barbusse, 93150 Le Blanc Mesnil, France. Tel: +33 1 48670257. Fax: +33 1 48670360. e-mail: firstname.lastname@example.org