Noncontact laser microsurgery of three-dimensional living objects for use in reproductive and regenerative medicine

Abstract

Abstract. Laser microsurgery has enabled us to make highly precise and delicate processing of living biological specimens. We present the results of using femtosecond (fs) laser pulses in assisted reproductive technologies. Femtosecond laser dissection of outer shells of embryos (so-called laser-assisted hatching) as well as laser-mediated detachment of the desired amount of trophectoderm cells (so-called embryo biopsy) required for preimplantaion genetic diagnosis were successfully performed. The parameters of laser radiation were optimized so as to efficiently perform embryo biopsy and preserve the viability of the treated embryos. Effects of application of fs-laser radiation in the infrared (1028 nm) and visible (514 nm) wavelength ranges were studied. We also applied laser microsurgery to develop a new simple reproducible model for studying repair and regeneration in vitro. Nanosecond laser pulses were applied to perform localized microdissection of cell spheroids. After microdissection, the edges of the wound surface opened, the destruction of the initial spheroid structure was observed in the wound area, with surviving cells changing their shape into a round one. It was shown that the spheroid form partially restored in the first six hours with subsequent complete restoration within seven days due to remodeling of surviving cells.

Genetic Material Manipulation and Modification by Optical Trapping and Nanosurgery-A Perspective

“…The method relies on glass micropipettes and a robotized stage to achieve the transfer. However, given the increasing spatial-temporal control in LS-OT setups (Yang et al, 2017;Sitnikov et al, 2018), similar results should be seen in the near future employing these setups. This successful methodology for mitochondrial transfer can have wide implications for the profiling and therapeutics of a whole family of diseases related to dysfunctional mitochondria (e.g., Huntington's disease, mitochondrial myopathies, mtDNA depletion syndrome, or cancer).…”