Evaluation of chemiluminescence and flow cytometry as tools in assessing production of hydrogen peroxide and superoxide anion in human spermatozoa Reda.

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Presentation transcript:

Evaluation of chemiluminescence and flow cytometry as tools in assessing production of hydrogen peroxide and superoxide anion in human spermatozoa Reda Mahfouz, M.D., Rakesh Sharma, Ph.D., Jakob Lackner, M.D., Nabil Aziz, M.D., Ashok Agarwal, Ph.D., H.C.L.D. Fertility and Sterility Volume 92, Issue 2, Pages 819-827 (August 2009) DOI: 10.1016/j.fertnstert.2008.05.087 Copyright © 2009 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Comparison of semen specimen neat, immature, and mature sperm before and after stimulation. (A) Intracellular H2O2: percentage of DCF+ve spermatozoa and (B) intracellular O2-•: percentage of HE+ve sperm. Fertility and Sterility 2009 92, 819-827DOI: (10.1016/j.fertnstert.2008.05.087) Copyright © 2009 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Dot plot histograms showing simultaneous measurements of intracellular hydrogen peroxide (upper panel) peroxide and intracellular superoxide levels (lower panel) in neat, immature, and mature spermatozoa. For hydrogen peroxide, the lower left quadrant represents viable, nonstained sperm, while the lower right shows viable sperm with high intracellular H2O2. The upper left quadrant shows apoptotic sperm. The upper right shows apoptotic sperm with high intracellular H2O2. For superoxide, the lower left quadrant represents viable, nonstained sperm, while the lower right shows apoptotic sperm. The upper left quadrant shows viable sperm with high intracellular O2−•, and the upper right shows apoptotic sperm with high intracellular O2−•. The numbers in parentheses represent the sperm population in each quadrant. Fertility and Sterility 2009 92, 819-827DOI: (10.1016/j.fertnstert.2008.05.087) Copyright © 2009 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Representative flow cytometry histograms showing the intracellular ROS levels measured by DCF for H2O2 and HE for O2−• in both chemiluminescence negative and positive specimens. A and B: luminol negative and positive samples; C and D: lucigenin negative and positive samples. Fertility and Sterility 2009 92, 819-827DOI: (10.1016/j.fertnstert.2008.05.087) Copyright © 2009 American Society for Reproductive Medicine Terms and Conditions