Abstract
Femtosecond laser pulses were applied for precise alphanumeric code engraving on the zona pellucida (ZP) of mouse zygotes for individual embryo marking and their identification. The optimal range of laser pulse energies required for safe ZP microsurgery has been determined. ZP was marked with codes in three different planes to simplify the process of embryo identification. No decrease in developmental rates and no morphological changes of embryos post laser-assisted engraving have been observed. ZP thickness of embryos post laser-assisted code engraving has been shown to differ significantly from that of control group embryos at the hatching stage. Due to moderate ZP thinning as compared to its initial width at 0.5 dpc (days post coitum), readability of the code degrades slightly and it still remains recognizable even at hatching stage. Our results demonstrate that application of femtosecond laser radiation could be an effective approach for noninvasive direct embryo tagging, enabling embryo identification for the whole period of preimplantation development.
Highlights
Lasers have become an efficient tool in assisted reproductive technologies (ART) [1,2,3]
Earlier studies of Sato et al [7] and Lenzi [8] have demonstrated the stimulating effect of red (λ = 647 nm) and infrared laser light on sperm motility
Openings in the outer shell surrounding oocytes and embryos required for sperm insertion during in vitro fertilization [12,15] or cell extraction during embryo biopsy can be created by means of lasers [13]
Summary
Lasers have become an efficient tool in assisted reproductive technologies (ART) [1,2,3]. Nowadays, infrared diode lasers (λ = 1.48 um) with milli- to microsecond pulse durations are the most popular lasers applied in the field of assisted reproduction for microdissection. Such systems are widely used for opening the ZP in assisted hatching [16,17,18,19,20]. The technique is based on femtosecond laser microsurgery of ZP and “engraving” small (~5 μm in width and ~20 μm in length) alphanumeric codes in the depth of ZP This technique may be useful in assisted reproductive technologies for preventing medical accidents relating to mixups. Our observations allow us to suppose that the technique proposed may be used for embryo labelling and for stimulating embryo hatching to start at prescribed location
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