Gold nanoparticle mediated cell manipulation using fs and ps laser pulses for cell perforation and transfection

Abstract

Manipulation of cells requires the delivery of membrane-impermeable substances like genetic materials or proteins into the cytoplasm. Thus delivery of molecules over the cell membrane barrier is one of the key technologies in molecular biology. Many techniques concerning especially the delivery foreign DNA have been developed. Notwithstanding there still is a range of applications where these standard techniques fail to raise the desired results due to low efficiencies, high toxicity or other safety issues. Especially the transfection of sensitive cell types like primary and stem cells can be problematic. Here we present an alternative, laser based technique to perforate the cell membrane and thus allowing efficient delivery of extra cellular molecules: Gold nanoparticles (GNP) are brought into close contact with the cell, were the laser-GNP interaction leads to membrane perforation. This allows the utilisation of a weakly focused laser beam leading to fast scanning of the sample and thus to a high throughput. To investigate the GNP-laser interaction in more detail we have compared membrane perforation obtained by different laser pulse lengths. From our results we assume strong light absorption for ps laser pulses and relatively small particles as the initiating perforation mechanism, whereas an enhanced near field scattering occurs at 200 nm GNP when using fs laser pulses. SEM and ESEM imaging were applied to give a deeper insight in the GNP-cell interaction and the effects of laser radiation on the GNP. Additionally dextran- FITC derivatives of varying sizes were used to investigate the impact of molecule size on delivery efficiency.