Intracytoplasmic sperm injection (ICSI) is the method of choice for the in vitro production (IVP) of equine embryos. However, conventional ICSI has been associated with mechanical damage to the oocyte caused by the deformation of the zona pellucida (ZP) and exposure of the oolemma to negative pressure during injection. Introduction of the less traumatic and more efficient piezo drill-assisted ICSI (PDAI) yielded higher cleavage rates and more consistent results. Nevertheless, PDAI is also associated with disadvantages such as the use of mercury and possible DNA damage. This led us to explore an alternative method avoiding oocyte trauma, namely laser-assisted ICSI (LAI), which involves creating a hole in the ZP prior to ICSI. In this pilot study, PDAI and LAI were compared for ICSI in the horse. No significant influences on subsequent embryonic development were observed.