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(a) Bessel beam transfection apparatus. A laser beam is demagnified and is incident upon an axicon which generates the Bessel beam. A telescope demagnifying by a factor of 8, consisting of an achromat and an aspheric lens, is placed in the beam path in order to create a Bessel beam with the optimum parameters for cell transfection. (b) Image of the generated Bessel beam incident upon the sample.
(a) Bessel beam “focus” is positioned on the cell plane. (b) Upon successful transfection, the cells express the red fluorescent protein and fluoresce red. (c) Bessel beam transfection vs Gaussian beam transfection. We achieve Bessel beam transfection of CHO cells for up to along the propagation axis of the beam. Each data point corresponds to the average transfection efficiencies obtained at a specific axial position and includes the number of spontaneously transfected cells, which varies between 0 and 5 for each sample dish. The magnified image of the area around can be viewed on the top right part of the main graph. As can be seen, the successful transfection of CHO cells can occur over 20 times the axial distance when using a Bessel beam, as compared to the Gaussian beam (when considering the threshold of efficiency to be 20%).
(a) Bessel beam passes through the obstructive layer of microspheres and suffers distortion. Due to the self-healing property of the Bessel beam, at away from the layer the beam profile is reformed and reaches the cell plane where photoporation will occur. (b) The Bessel beam profile is distorted after passing through the microsphere obstructive layer. (c) The Bessel beam is reconstructed after (depending on the sample) subsequently imaged on the cell membrane and photoporation takes place.
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