Single point single-cell nanoparticle mediated pulsed laser optoporation.

ABSTRACT

This article presents an optical platform for studying the dynamics of nanoparticle assisted pulsed laser optoporation of individual living cells. Here plasmonic nanoparticles (NPs) act as markers of the exact spatial position of living cell membranes and as an enhancer for localized pulsed laser perforation. High contrast NP imaging using reflected light microscopy (RLM) allows accurate and automatic laser targeting at individual NPs for spatially controlled laser optoporation of single cells at a single point. The NP imaging method is compatible with fluorescence microscopy and a cellular incubator that allows study of real-time perforation kinetics of live cells and the optomechanical interaction of NPs with membranes. These parameters are of great interest for the development and experimental implementation of the technology of pulsed laser optoporation and transfection applied to single living cells as well as to bulk-level assays.