Ultrafast lasers, with pulse durations below a few picoseconds, are of significant interest to the industry, offering a cutting-edge approach to enhancing manufacturing processes and enabling the fabrication of intricate components with unparalleled accuracy. When processing metals at irradiances exceeding the evaporation threshold of about 1010 W/cm² these processes can generate ultra-short, soft X-ray pulses with photon energies above 5 keV. This has prompted extensive discussions and regulatory measures on radiation safety. However, the impact of these ultra-short X-ray pulses on molecular pathways in the context of living cells, has not been investigated so far. This paper presents the first molecular characterization of epithelial cell responses to ultra-short soft X-ray pulses, generated during processing of steel with an ultrafast laser. The laser provided pulses of 6.7 ps with a pulse repetition rate of 300 kHz and an average power of 500 W. The irradiance was 1.95 ×1013 W/cm2. Ambient exposure of vitro human cell cultures, followed by imaging of the DNA damage response and fitting of the data to a calibrated model for the absorbed dose, revealed a linear increase in the DNA damage response relative to the exposure dose. This is in line with findings from work using continuous wave soft X-ray sources and suggests that the ultra-short X-ray pulses do not generate additional hazard. This research contributes valuable insights into the biological effects of ultrafast laser processes and their potential implications for user safety.
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