Femtosecond laser micro machining of surfaces knows a gain of interest as it demonstrates efficient and precise processing with reduced side effects around the irradiated zone, and also because of the remarkable costs reduction and reliability improvements of nowadays commercially available sources. Controlling the intensity distribution spatially can offer a supplementary degree of flexibility and precision in achieving user-defined ablation spatial profile, drilling, cutting of materials or in-volume laser-induced modifications. In this scope, the possibility to generate a top-hat intensity distribution by spatially shaping the beam wavefront is studied in this work. An optimization of Zernike polynomials coefficients is conducted to numerically determine an adequate phase mask that shapes the laser intensity distribution following a targeted top hat distribution in the processing plane, usually at the focal length of a converging lens. The efficiency of the method is numerically investigated in the optimization by evaluation of the root mean square error (RMS) between the top-hat target and the calculated laser distribution in the far field. We numerically verify that acceptable top-hat beam shaping of various size can be achieved with a sufficient number of Zernike polynomials, opening the way to phase mask calculations adapted to the wavefront modulator ability to reproduce Zernike polynomials.
Corneal therapeutic molecules delivery represents a promising solution to maintain human corneal endothelial cells (HCECs) viability, but the difficulty is transport across cell membrane. A new delivery method published recently consists in ephemerally permeabilizing cell membranes using a photo-acoustic reaction produced by carbon nanoparticles (CNPs) and femtosecond laser (FsL). The aim of this work is to investigate the size of pores formed at cell membrane by this technique. To induce cell permeabilization, HCECs were put in contact with CNPs and irradiated with a 500 μm diameter Ti:Sa FsL focalized spot. Four sizes of marker molecules were delivered into HCECs to investigate pore sizes: calcein (1.2 nm), FITC-Dextran 4kDa (2.8 nm) and FITC-Dextran 70kDa (12 nm) and FITC-Dextran 2MDa (50 nm). Delivery of each molecule was assessed by flow cytometry, a technique able to measure their presence into cells. We showed that the delivery rate was dependent of their size. Calcein was delivered in 56.1±8.2% of HCECs, FITC-Dextran 4kDa in 42.2±3.5%, FITC-Dextran 70 kDa in 21.5±2.7% and finally FITC-Dextran 2MDa in 12.9±2.0%. It means that a large number of pores in the size ranging from 1.2 to 2.8 nm were formed. However, 12 nm and larger pores were almost half more infrequent. Pore sizes formed at cell membrane by the technique of cell permeabilization by FsL activated CNPs was investigated. The results indicated that the pore sizes are large enough for the efficient delivery of small, medium and big therapeutics molecules on HCECs by this technique.