Diagnostic accuracy of sperm chromatin dispersion test to evaluate sperm deoxyribonucleic acid damage in men with unexplained infertility Cinthia M.

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Diagnostic accuracy of sperm chromatin dispersion test to evaluate sperm deoxyribonucleic acid damage in men with unexplained infertility Cinthia M. Feijó, M.Sc., Sandro C. Esteves, M.D., Ph.D. Fertility and Sterility Volume 101, Issue 1, Pages 58-63.e3 (January 2014) DOI: 10.1016/j.fertnstert.2013.09.002 Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Relationship between the SCD test and the TUNEL assay for assessing sperm DNA damage in ejaculates of 20 men with unexplained infertility. Fertility and Sterility 2014 101, 58-63.e3DOI: (10.1016/j.fertnstert.2013.09.002) Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Receiver operator characteristic curves showing four different TUNEL thresholds (4%, 10%, 12%, and 19%) to select the optimal cutoff points of SCD to discriminate between patients with normal and abnormal levels of sperm DNA damage. The SCD cutoff points yielding the largest AUC for each TUNEL threshold are presented (arrows). Fertility and Sterility 2014 101, 58-63.e3DOI: (10.1016/j.fertnstert.2013.09.002) Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

Supplemental Figure 1 Assessment of DNA damage in sperm. (A) TUNEL assay. Sperm showing bright apple-green fluorescence represent cells with damaged DNA (red arrows), in which dUTP is incorporated to DNA breaks, in contrast to non–DNA-damaged sperm (white arrows) that have been first identified using DAPI staining (not shown). Sperm photomicrographs obtained at ×1,000 using a fluorescence microscope (Eclipse E600; Nikon) equipped with an epi-illumination module and a mercury ultraviolet source. Filter B2A was used for FITC-dUTP. (B) SCD test. Nucleoids obtained with the improved SCD procedure (Halosperm; Halotech DNA) under bright-field microscopy and Wright's stain. The red arrow indicates a spermatozoon containing a normal DNA molecule (a halo of dispersed chromatin is seen). The blue arrow indicates a spermatozoon with damaged DNA molecule (an absent halo of dispersed chromatin is seen). Microphotographs obtained at ×1,000 using bright-field microscopy (Alphaphot-2 YS; Nikon). Fertility and Sterility 2014 101, 58-63.e3DOI: (10.1016/j.fertnstert.2013.09.002) Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions