Zygote

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  • Copyright © Cambridge University Press 2014 The online version of this article is published within an Open Access environment subject to the conditions of the Creative Commons Attribution-NonCommercial-ShareAlike licence <
  • DOI: http://dx.doi.org/10.1017/S0967199414000288 (About DOI), 7 pages. Published online: 03 July 2014
  • OPEN ACCESS

A double-blinded comparison of in situ TUNEL and aniline blue versus flow cytometry acridine orange for the determination of sperm DNA fragmentation and nucleus decondensation state index

Jamal Hamidia2, Christophe Frainaisa2, Edouard Amara3, Eric Baillya4, Patrice Clémenta2 and Yves Ménézoa1a3a4a5 c1

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.

Summary

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)

Keywords:

  • Acridine orange;
  • Aniline blue;
  • Decondensation;
  • DNA fragmentation;
  • Flow cytometry;
  • Sperm;
  • TUNEL

Correspondence

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: yves.menezo@gmail.com

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