Mouse sperm sorted using space-constrained microfluidic sorting (SCMS) system with 15 mm long channel was compared to those using swim-up technique and non-sorted sperm. The comparisons were performed for (A) Curvilinear velocity, (B) Straight-line velocity, (C) Linearity, and (D) percentage of motile sperm for 30 min of incubation. SCMS system with 15 mm length and 30 min of incubation resulted in sperm with higher motility and percentage of motile sperm compared to swim-up technique and non-sorted sperm. Data were presented as average ± standard error (SEM) (N=4–7). (E) The sperm curvilinear velocity, straight-line velocity, and collectable sperm percentage were compared after sorting using space-constrained microfluidic sorting (SCMS) system with different channel lengths (7 mm, 10 mm, 15 mm and 20 mm) for 30 minutes of incubation. Data were presented as average ± standard error (SEM) (N=3).