Flow Cytometric Methods for Sperm Assessment

  • Vanesa Robles
  • Felipe Martínez-Pastor
Part of the Methods in Molecular Biology book series (MIMB, volume 927)


Flow cytometry allows the assessment of multiple sperm parameters following a diverse number of protocols. Here, we describe three methods for evaluating critical aspects of sperm quality: the double stain SYBR-14 and propidium iodide (PI) for assessing sperm viability; the double stain PNA-FITC (peanut agglutinin-fluorescein isothiocyanate) and PI for assessing acrosomal integrity combined with sperm viability; and JC-1 (5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethyl-benzimidazolylcarbocyanine iodide) for assessing mitochondrial membrane potential. These three stains are widely used and can be analyzed with basic flow cytometers.

Key words

Flow cytometry Spermatozoa Viability Mitochondria Acrosome Propidium iodide SYBR-14 JC-1 PNA FITC 



This work was supported in part by grants LE019A10-2 (Junta de Castilla y León), AGL2010-15758 (MICINN) and Fundación Ramón Areces. The authors were supported by the Ramón y Cajal program (MICINN). The authors thank María Mata-Campuzano for producing several of the microphotographs and cytograms.


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Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  1. 1.INDEGSALUniversity of LeónLeónSpain

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