7 March 2014 Femtosecond optical injection of intact plant cells using a reconfigurable platform
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Abstract
The use of ultrashort-pulsed lasers for molecule delivery and transfection has proved to be a non-invasive and highly efficient technique for a wide range of mammalian cells. This present study investigates the effectiveness of femtosecond photoporation in plant cells, a hard-to-manipulate yet agriculturally relevant cell type, specifically suspension tobacco BY-2 cells. Both spatial and temporal shaping of the light field is employed to optimise the delivery of membrane impermeable molecules into plant cells using a reconfigurable optical system designed to be able to switch easily between different spatial modes and pulse durations. The use of a propagation invariant Bessel beam was found to increase the number of cells that could be viably optoinjected, when compared to the use of a Gaussian beam. Photoporation with a laser producing sub-12 fs pulses, coupled with a dispersion compensation system to retain the pulse duration at focus, reduced the power required for efficient optical injection by 1.5-1.8 times when compared to a photoporation with a 140 fs laser output.
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Claire A. Mitchell, Claire A. Mitchell, Stefan Kalies, Stefan Kalies, Tomas Cizmar, Tomas Cizmar, Nicola Bellini, Nicola Bellini, Anisha Kubasik-Thayil, Anisha Kubasik-Thayil, Alexander Heisterkamp, Alexander Heisterkamp, Lesley Torrance, Lesley Torrance, Alison G. Roberts, Alison G. Roberts, Frank J. Gunn-Moore, Frank J. Gunn-Moore, Kishan Dholakia, Kishan Dholakia, } "Femtosecond optical injection of intact plant cells using a reconfigurable platform", Proc. SPIE 8972, Frontiers in Ultrafast Optics: Biomedical, Scientific, and Industrial Applications XIV, 89720C (7 March 2014); doi: 10.1117/12.2037784; https://doi.org/10.1117/12.2037784
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