Molecule transfer into mammalian cells by single sub-nanosecond laser pulses.
Florian HausladenPetra KruseFelicia HessenbergerThomas StegmayerYu-Ting KaoWolf SeelertRosemarie PreyerMarco SpringerKarl StockRainer WittigPublished in: Journal of biophotonics (2023)
A rapid, precise, and viability-retaining method for cytoplasmic molecule delivery is highly desired for cell engineering. Routine methods suffer from low throughput, lack of selectivity, requirement of helper compounds, predominant endosomal delivery, and/or are restricted to specific molecule classes. Photonic cell manipulation bears potential to overcome these drawbacks. Here we investigated mammalian cell manipulation by single sub-nanosecond laser pulses. Axial beam waist positioning close to a cell monolayer induced culture vessel damage and zones of cell ablation. Cells at margins of ablation zones exhibited uptake of membrane-impermeant fluorophores and GFP expression plasmids. Increasing Rayleigh-length and beam waist diameter reduced the sensitivity to axial defocusing and resulted in robust molecule transfer. Serial application of single pulses focused over a moving cell monolayer yielded quantitative molecule transfer to cells at rates up to 40%. Our results could be basic to spatially and temporally controlled single laser pulse-mediated marker-free high throughput cell manipulation. This article is protected by copyright. All rights reserved.
Keyphrases
- single cell
- cell therapy
- high throughput
- escherichia coli
- body mass index
- stem cells
- induced apoptosis
- poor prognosis
- mass spectrometry
- mesenchymal stem cells
- cell death
- dendritic cells
- atrial fibrillation
- long non coding rna
- climate change
- cell cycle arrest
- immune response
- atomic force microscopy
- binding protein
- multidrug resistant
- radiofrequency ablation