For the first time, the data have been obtained on the effects of high-intensity terahertz (THz) radiation (with the intensity of 30 GW/cm2, electric field strength of 3.5 MV/cm) on human skin fibroblasts. A quantitative estimation of the number of histone Н2АХ foci of phosphorylation was performed. The number of foci per cell was studied depending on the irradiation time, as well as on the THz pulse energy. The performed studies have shown that the appearance of the foci is not related to either the oxidative stress (the cells preserve their morphology, cytoskeleton structure, and the reactive oxygen species content does not exceed the control values), or the thermal effect of THz radiation. The prolonged irradiation of fibroblasts also did not result in a decrease of their proliferative index.

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Purpose Our purpose was to study whether application of femtosecond laser pulses for alphanumeric code marking in the volume of zona pellucida (ZP) could be effective and reliable approach for direct tagging of preimplantation embryos. Methods Femtosecond laser pulses (wavelength of 514 nm, pulse duration of 280 fs, repetition rate of 2.5 kHz, pulse energy of 20 nJ) were applied for precise alphanumeric code engraving on the ZP of mouse embryos at the zygote stage for individual embryo marking and their accurate identification. Embryo quality assessment every 24 h post laser-assisted marking as well as immunofluorescence staining (for ICM/TE cell number ratio calculation) were performed. Results Initial experiments have started with embryo marking in a single equatorial plane. The codes engraved could be clearly recognized until the thinning of the ZP prior to hatching. Since embryo may change its orientation during the ART cycle, multiplane code engraving seems to be more practical for simplifying the process of code searching and embryo identification. We have marked the ZP in three planes, and no decrease in developmental rates as well as no morphological changes of embryos post laser-assisted engraving have been observed as compared to control group embryos. Conclusions Our results demonstrate the suitability of femtosecond laser as a novel tool for noninvasive embryo tagging, enabling embryo identification from day 0.5 post coitum to at least early blastocyst stage. Thus, the versatility and the potential use of femtosecond lasers in the field of developmental biology and assisted reproduction have been shown.

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