Near-infrared laser cellular ablation and development in Xenopus laevis embryos

Abstract

The biological applications of pulsed near-infrared (NIR) lasers are widening due to potential use of this technology to manipulate cellular structure. The present study was conducted to further explore the efficacy of NIR lasers for ablating individual cells in eight-cell stage Xenopus laevis embryos. Ablations were performed with four experimental groups, at magnifications of 10× and 40× (laser beam radius of 0.32 and 0.23 μm, respectively) with either one cell or two adjacent cells targeted. The survivorship, size, and phenotypic mutations of each group were documented and compared to a control group. Survivorship was not affected in any experimental group; however, statistically significant differences were noted in embryonic length (p = 0.02) and in morphology (p < 0.01) for the experimental groups with ablation of two cells at 40× (laser beam radius of 0.23 μm) magnification. The survivorship of targeted embryos in this experiment encourages the use of NIR radiation for cell ablation studies. Our study demonstrated that NIR laser spectroscopy could complement existing X. laevis fate mapping data and improve understanding of developmental plasticity in early embryos as well as serve as a clinical tool for removing pathological cells. Despite visible effects on embryo development, the damages incurred in the most extreme condition did not significantly impact survivorship through the early tadpole stage. This finding does not rule out latent effects that might affect fitness of older tadpoles or even adult frogs.