Objective To study a new sorting method for motile human sperm using passively polydimethylsiloxane (PDMS) microfluidic chip.Methods 40 sperm specimens are separated by the PDMS microfluidic chip sorting device,which was home-made,and the semen parameters such as sperm motility,morphology and hypoosmotic swelling rate were compared before and after the experiment.Meanwhile,thirty motile sperm samples were prepared by density gradient centrifugal separation in order to assess sperm motility using PDMS micro-fluidic chip.Results There was significant difference between pre-sorting and post-sorting in terms of sperm motility,morphology and hypoosmotic swelling rate.Sperm motility increased from (29.4 ± 8.1) % to (94.2 ± 3.3) %(t=24.58,P<0.01),the normal morphology sperm increased from (10.5 ± 8.0) % to (30.0± 5.5) % (t=13.67,P < 0.01) and hypoosmotic swelling rate increased from (65.5 ± 7.4) % to (96.0 ± 1.8) % (t=10.13,P < 0.01).There was no difference in sperm motility between spermatozoa with and without exposure to PDMS ((71 ± 3.4) % / (70 ± 2.5) %,P>0.05).Conclusions The microfluidic chip sorting device provides a novel method for sperm motile sorting,morphologically normal spermatozoa from semen without centrifugation.This technology may be useful in sorting motile spermatozoa and insemination and in vitro fertilization to improve birth health quality.