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Hum. Reprod. Advance Access originally published online on July 15, 2004
Human Reproduction 2004 19(10):2267-2276; doi:10.1093/humrep/deh416
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Human Reproduction vol. 19 no. 10 © European Society of Human Reproduction and Embryology 2004; all rights reserved

Comparison of four fluorochromes for the detection of the inner mitochondrial membrane potential in human spermatozoa and their correlation with sperm motility

Carole Marchetti1,2,3, Nathalie Jouy4, Brigitte Leroy-Martin2,3, André Defossez2,3, Pierre Formstecher1 and Philippe Marchetti1,5

1 INSERM U459, 3 Laboratoire d'Histologie, 4 IFR 114-IMPRT, Faculté de Médecine 1, Place Verdun, 59045 Lille Cedex and 2 Laboratoire de Biologie de la Reproduction, Hôpital Jeanne de Flandre, 59037 Lille Cedex, France

5 To whom correspondence should be addressed. Email: philippe.marchetti{at}lille.inserm.fr

BACKGROUND: Sperm motility evaluation is associated with fertility in IVF programmes. The visual estimation of sperm motility is extremely subjective. Hence, alternative methods are required. Among them, determination of mitochondrial membrane potential ({Delta}{psi}m) changes of spermatozoa using potentiometric dyes may be a reliable test to determine sperm quality. However, the use of the potentiometric dyes in sperm samples has not been compared. METHODS: We have studied sperm samples from 28 infertile patients enrolled in an IVF programme in flow cytometry after staining of spermatozoa with four commonly used potentiometric dyes. Sperm motility was evaluated visually. RESULTS: As expected, JC-1 seems to detect specifically {Delta}{psi}m changes, CMX-Ros, DiOC6(3) and TMRE fluorescence is easily analysed and the latter three fluorochromes are particularly suitable for multiparametric staining. Irrespective of the {Delta}{psi}m-dependent fluorochromes used to stain spermatozoa, a positive correlation was found between the percentage of {Delta}{psi}mhigh cells and forward motility and also with high fertilization rates after IVF. CONCLUSION: The four fluorochromes may be useful for evaluation of sperm samples from infertile patients. The choice of the potentiometric dyes will depend on their fluorescence characteristics in order to use them in combination with other fluorescent markers.

Key words: flow cytometry/IVF/mitochondria/spermatozoa


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