Fluorimetric techniques for the assessment of sperm membranes

Alisa Komsky-Elbaz*, Zvi Roth

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

8 Scopus citations


Standard spermiograms describing sperm quality are mostly based on the physiological and visual parameters, such as ejaculate volume and concentration, motility and progressive motility, and sperm morphology and viability. However, none of these assessments is good enough to predict the semen quality. Given that maintenance of sperm viability and fertilization potential depends on membrane integrity and intracellular functionality, evaluation of these parameters might enable a better prediction of sperm fertilization competence. Here, we describe three feasible methods to evaluate sperm quality using specific fluorescent probes combined with fluorescence microscopy or flow cytometry analyses. Analyses assessed plasma membrane integrity using 4',6-diamidino-2-phenylindole (DAPI) and propidium iodide (PI), acrosomal membrane integrity using fluorescein isothiocyanate-conjugated Pisum sativum agglutinin (FITC-PSA) and mitochondrial membrane integrity using 5,5',6,6'-tetra-chloro-1,1',3,3'-tetraethylbenzimidazolyl carbocyanine iodide (JC-1). Combinations of these methods are also presented. For instance, use of annexin V combined with PI fluorochromes enables assessing apoptosis and calculating the proportion of apoptotic sperm (apoptotic index). We believe that these methodologies, which are based on examining spermatozoon membranes, are very useful for the evaluation of sperm quality.

Original languageAmerican English
Article numbere58622
JournalJournal of Visualized Experiments
Issue number141
StatePublished - Nov 2018

Bibliographical note

Publisher Copyright:
© 2018 Journal of Visualized Experiments.


  • Acrosome reaction
  • Developmental biology
  • Flow cytometry
  • Fluorescent probe
  • Issue 141
  • Mitochondrial membrane potential
  • Plasma membrane
  • Semen evaluation
  • Sperm


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