Objective: To directly compare distinct assays proposed to monitor human sperm quality and possibly preselect sperm populations for assisted reproductive technology (ART). Design: Analysis of human sperm sample quality using several methodologies. Setting: Academic and clinical institutions. Patient(s): Samples from consenting patients undergoing routine semen analysis or ART. Interventions: Human sperm samples were analyzed in terms of World Health Organization parameters and processed for annexin V, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling of DNA (TUNEL), and the sperm-ubiquitin tag immunoassay (SUTI). Samples were analyzed both by flow cytometry and fluorescence microscopy. Main Outcome Measure(s): Correlations among apoptotic markers (outer leaflet phosphatidylserine exposure, membrane integrity, and DNA fragmentation), external ubiquitination, and semen parameters in human spermatozoa. Result(s):Nonviable sperm, TUNEL-positive cells, and ubiquitin fluorescence intensity means inversely correlate with semen parameters. Apoptotic markers do not correlate with sperm surface ubiquitination. Normozoospermic samples have a higher number of viable cells and lower DNA fragmentation compared with samples with abnormal parameters. Nonviable sperm are more prevalent in samples with low counts and poor morphology but not low motility. Not all sperm with morphologic abnormalities present surface ubiquitination. Conclusion(s):Sperm quality is inversely correlated with lack of viability, DNA fragmentation, and ubiquitin fluorescence intensity means. However, none of the apoptotic markers correlate with ubiquitin labeling. Elimination of defective sperm cells prior to ART using surface markers (annexin V, ubiquitin) seems unwarranted at this stage. (Fertil Steril 2007;87:572-83.

Retention of the second polar body and chromosome malsegregation were observed after parthenogenetic activation, either spontaneous or induced by puromycin. This means that using parthenogenetic embryos for stem cell research will require great care and attention.

This is the first description of a simple and fast hybridization protocol for spermatozoa without a decondensation step, allowing preservation of the morphology of the sperm head that is particularly useful to correlate abnormal spermatozoa with specific chromosome aneuploidies. With this technique we were able to avoid troubles in interpretation of FISH spots that does not depend on the quality of nuclear decondensation, as it is the case in the previously described methods. Our goal was to demonstrate the efficiency of the method without loosing sperm head morphology. Further studies are needed to correlate the aneuploidy rates for specific chromosomes with sperm morphology.