12 February 2009 Cell damage extent due to irradiation with nanosecond laser pulses under cell culturing medium and dry environment
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Abstract
Cell mono-layers were irradiated with nanosecond laser pulses under two distinct scenarios: (a) with culturing medium positioning the beam waist at different stand-off distances γ and (b) without cell culturing medium, positioning the beam waist directly on top of the cell mono-layer. Damaged cells were marked with Trypan Blue, a vital cell marker. Three different zones of damage were identified: (1) a zone of complete cell clearance, surrounded by (2) a ring of dead cells marked with Trypan Blue and (3) the rest of the cell culture where the cells remain alive and viable. Different hydrodynamic mechanisms damage cells as it was shown by high speed video for γ=0 and comparison with time resolved imaging. The cell damage mechanism has its origin on the optical breakdown plasma formation. For the case with culturing medium, a combination of plasma formation and shear stresses are responsible for cell damage; wheras for the case without cell culturing medium, the plasma formation is the only mechanism of interaction between laser pulses and cells. The rapidly expanding plasma generates shock waves whose pressure is most likely responsible for the cell detachment observed.
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Francisco G. Pérez-Gutiérrez, Francisco G. Pérez-Gutiérrez, Gabriel Guillen, Gabriel Guillen, Rodger Evans, Rodger Evans, Santiago Camacho-López, Santiago Camacho-López, Guillermo Aguilar, Guillermo Aguilar, } "Cell damage extent due to irradiation with nanosecond laser pulses under cell culturing medium and dry environment", Proc. SPIE 7175, Optical Interactions with Tissue and Cells XX, 717514 (12 February 2009); doi: 10.1117/12.816562; https://doi.org/10.1117/12.816562
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