Molecule transfer into mammalian cells by single sub‐nanosecond laser pulses

• DOI: 10.1002/jbio.202200327
• Publication Date: 2023-01-12T13:27:11Z
• Authors: Florian Hausladen; Petra Kruse; Felicia Hessenberger; Thomas Stegmayer; Yu‐Ting Kao; Wolf Seelert; Rosemarie Preyer; Marco Springer; Karl Stock; Rainer Wittig
• Keywords: Nanosecond

A rapid, precise, and viability-retaining method for cytoplasmic molecule delivery is highly desired cell engineering. Routine methods suffer from low throughput, lack of selectivity, requirement helper compounds, predominant endosomal delivery, and/or are restricted to specific classes. Photonic manipulation bears the potential overcome these drawbacks. Here we investigated mammalian by single sub-nanosecond laser pulses. Axial beam waist positioning close a monolayer induced culture vessel damage zones ablation. Cells at margins ablation exhibited uptake membrane-impermeant fluorophores GFP expression plasmids. Increasing Rayleigh-length diameter reduced sensitivity axial defocusing resulted in robust transfer. Serial application pulses focused over moving yielded quantitative transfer cells rates up 40%. Our results could be basic spatially temporally controlled pulse-mediated marker-free high throughput manipulation.