title: 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.
abstract: 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.
authors.affiliation: Laboratoire Clément,17 avenue d'Eylau,75016 Paris and 8 avenue Henri Barbusse,93150 Le Blanc Mesnil,France.
authors.forename: Jamal
authors.initials: J
authors.lastname: Hamidi
authors.affiliation: Laboratoire Clément,17 avenue d'Eylau,75016 Paris and 8 avenue Henri Barbusse,93150 Le Blanc Mesnil,France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Frainais
authors.affiliation: Urology Department,American Hopsital of Paris,92200 Neuilly-sur-Seine,France.
authors.forename: Edouard
authors.initials: E
authors.lastname: Amar
authors.affiliation: Beckman Coulter France,Villepinte,95942 Roissy,France.
authors.forename: Eric
authors.initials: E
authors.lastname: Bailly
authors.affiliation: Laboratoire Clément,17 avenue d'Eylau,75016 Paris and 8 avenue Henri Barbusse,93150 Le Blanc Mesnil,France.
authors.forename: Patrice
authors.initials: P
authors.lastname: Clément
authors.affiliation: Yves Ménézo. Laboratoire Clément,17 avenue d'Eylau,75016 Paris and 8 avenue Henri Barbusse,93150 Le Blanc Mesnil,France.
authors.forename: Yves
authors.initials: Y
authors.lastname: Ménézo
mesh_terms: D000165:Acridine Orange; D000814:Aniline Compounds; D053938:DNA Fragmentation; D004311:Double-Blind Method; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes; D006801:Humans; D020287:In Situ Nick-End Labeling / Q000379:methods*; D008297:Male; D027724:Reproductive Techniques, Assisted; D013094:Spermatozoa / Q000166:cytology / Q000502:physiology*
chemical_list: D000814:Aniline Compounds; D005456:Fluorescent Dyes; C017006:aniline blue; D000165:Acridine Orange
keywords: Acridine orange; Aniline blue; DNA fragmentation; Decondensation; Flow cytometry; Sperm; TUNEL