High-throughput optical injection of mammalian cells using a Bessel light beam

Femtosecond photoporation is an optical method for the injection of membrane impermeable substances into cells. Typically this is a low-throughput method where each cell is individually targeted. Here, we present a novel microfluidic platform with passive optical injection improving previously reported throughputs by one order of magnitude. In this new geometry, two-dimensional hydrodynamic focusing is achieved using a three-dimensional nozzle which confines mammalian cells to the central region of the microfluidic channel. A reusable quartz chip is designed so that a propagation invariant, 'non-diffracting' Bessel beam can be directed along the centre of the channel, parallel to but counter-propagating with the flow of cells in contrast to previous orthogonal geometries. This allows for higher flow speeds to be used whilst maintaining the necessary dwell time for cells in the core of the Bessel beam. Using this method, we have achieved viable injection of HL60 cells with propidium iodide with an efficiency of 20.4 ± 4.2% and CHO-K1 cells (31.0 ± 9.5%) at a rate of up to 10 cells s^-1.