ROS formation was measured by chemiluminescence assay with luminol (5-amino-2,3-dihydro-1,4-phthalazinedione) as the probe, using an AutoLumat LB 953 multi-tube luminometer (Berthold Technologies, Oak Ridge, TN, USA). Results were expressed as relative light units RLU/s/106 sperm.25 (link) Total antioxidant capacity (TAC) was measured in the seminal plasma using the antioxidant assay kit (Cayman Chemical, Ann Arbor, MI, USA). Trolox standards and reagent were prepared as per the manufacturer's instructions at the time of the assay. Absorbance was monitored at 750 nm using ELx800 Absorbance Microplate Reader. Results were expressed as micromoles of Trolox.21 (link)
Sperm DNA fragmentation was evaluated using a terminal deoxynucleotidyl transferase–mediated fluorescein–dUTP nick end labeling (TUNEL) assay with an apoptosis-detection kit (Apo-Direct, BD Biosciences Pharmingen, San Diego, CA, USA) and flow cytometry. All fluorescence signals of labeled spermatozoa were analyzed by flow cytometer. Totally, 10 000 spermatozoa were examined, and the percentage of TUNEL-positive cells was calculated.26 (link)
Sperm DNA fragmentation was evaluated using a terminal deoxynucleotidyl transferase–mediated fluorescein–dUTP nick end labeling (TUNEL) assay with an apoptosis-detection kit (Apo-Direct, BD Biosciences Pharmingen, San Diego, CA, USA) and flow cytometry. All fluorescence signals of labeled spermatozoa were analyzed by flow cytometer. Totally, 10 000 spermatozoa were examined, and the percentage of TUNEL-positive cells was calculated.26 (link)
