Fig. 4.
Effects of pipetting on intracellular reactive oxygen species (ROS) levels in epididymal rat spermatozoa. (a) To evaluate the effect of pipetting on intracellular ROS in epididymal rat spermatozoa, DCF mean fluorescence intensity (MFI) was measured under basal (without tert-butyl hydroperoxide [TBHP] treatment) and stimulated (+TBHP) conditions in total and viable sperm populations that were subjected to pipetting. (b) In addition, the percentage of viable spermatozoa was evaluated under basal and stimulated conditions after pipetting of sperm. (c, d) To determine susceptibility to the ROS-stimulating product, fold increases in DCFMFI (c) and decreases in viability (d) following TBHP treatment were calculated. C, control; P, pipetting. Data are the mean ± s.e.m. (n = 8). *P < 0.05 compared with control. †P < 0.05 compared with basal ROS production.