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The aim of this study was to find the proper method for sperms immobilization without reducing DNA integrity and damaging the shape of bovine sperms . Sperm immobilization was carried out by double shots using laser Octax MTG with wavelength of 1.48 μm and Eye Ware 3.0 software. A total of 20 sperms were shoot at 4 points, which was the tail end, the mid-tail, the neck and the head of the sperms. The shooting of 1.48 μm laser had the highest effect on immobility reaching 100% on the head and the DNA integrity of the sperms reached up to 90%, the shape of sperms were remained normal (100%) and did not showing significant effect. The results of this study indicated that (1) the laser diode with a wavelength of 1.48 μm at the mid-tail section of the sperm was most effective for sperm immobilization; (2) the use of laser diodes with a wavelength of 1.48 μm can be used to immobilize the sperms before intracytoplasmic sperm injection (ICSI). The diode laser shooting with a wavelength of 1.48 μm at the tail was the most effective in immobilizing the sperms with an immobility of 95.00% ± 5.98 and DNA integrity of 85.00% ± 11.65.