Direct Methods for the Detection of Reactive Oxygen Species in Human Semen Samples

R. John Aitken, Geoffry N. De Iuliis, and Mark A. Baker

Abstract The accurate, selective measurement of reactive oxygen species (ROS) production by human spermatozoa is still a work-in-progress. Traditionally, chemi-luminescence approaches have been employed using luminol or lucigenin as probes to provide a sensitive readout of the redox status of human sperm populations. A particular concern with these chemiluminescent probes is that they can also generate spurious signals as a consequence of redox cycling activity, particularly in the case of lucigenin which only needs to experience a one-electron reduction at the hands of a cellular oxidoreductase to generate intense but artificial redox signals. Most importantly, the measurement of ROS in sperm suspensions is confounded by the presence of leukocytes. Alternatives are to either physically remove the leukocytes using magnetic beads coated with a monoclonal antibody against the common leukocyte antigen (CD45) or to use fluorescent probes in combination with fluorescence microscopy or flow cytometry to focus the analysis on individual cells rather than cell suspensions.

Keywords Chemiluminescence techniques • Direct methods • Detection of reactive oxygen species • Semen • Chemiluminescent assays

R.J. Aitken, PhD, ScD (*) • G.N. De Iuliis, PhD, BSc (Hons)

Department of Biological Sciences, University of Newcastle, Callaghan, NSW 2308, Australia e-mail: [email protected]

Department of Environmental and Life Science, University of Newcastle, Callaghan, NSW, Australia

A. Agarwal et al. (eds.), Studies on Men's Health and Fertility, Oxidative Stress 275

in Applied Basic Research and Clinical Practice, DOI 10.1007/978-1-61779-776-7_14, © Springer Science+Business Media, LLC 2012

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