Phototoxicity of brightfield live-cell imaging on murine ovarian follicles.

PURPOSE: Within the female reproductive tract, fertilization and embryo development occur in the complete absence of light. Light exposure during the in vitro manipulation of ovarian follicles negatively affects follicle viability. Laboratory conditions involve exposure to varying wavelengths, intensities, and light sources.

METHODS

In our study, we observed the effect of recurring light exposure on murine ovarian follicle viability subsequently assessed by evaluating increase in diameter, changes in morphology, and results from the LIVE/DEAD assay. A time-lapse approach allowed for effective observation of follicle interactions and viability, with no relevant movement or loss of focus throughout the imaging period.

RESULTS

However, follicles exposed to light showed a significant increase in granulosa cell death, as indicated by the LIVE/DEAD assay. No significant difference was observed in regard to morphology.

CONCLUSION

Brightfield time-lapse provides a valuable tool for identifying early predictive signs and key milestones in follicle development, which may enhance in vitro follicle culture techniques and improve patient treatment outcomes, but simultaneously carries a risk of phototoxicity that must be considered when applying this method.