Continuous medium exchange and optically induced electroporation of cells in an integrated microfluidic system

Optically induced electroporation(OIE)is a promising microfluidic-based approach for the electroporation of cell membranes.However,previously proposed microfluidic cell-electroporation devices required tedious sample pre-treatment steps,specifically,periodic media exchange.To enable the use of this OIE process in a practical protocol,we developed a new design for a microfluidic device that can perform continuous OIE;i.e.,it is capable of automatically replacing the culture medium with electroporation buffers.Integrating medium exchanges on-chip with OIE minimises critical issues such as cell loss and damage,both of which are common in traditional,centrifuge-based approaches.Most importantly,our new system is suitable for handling small or rare cell populations.Two medium exchange modules,including a micropost array railing structure and a deterministic lateral displacement structure,were first adopted and optimised for medium exchange and then integrated with the OIE module.The efficacy of these integrated microfluidic systems was demonstrated by transfecting an enhanced green fluorescent protein(EGFP)plasmid into human embryonic kidney 293T cells,with an efficiency of 8.3%.This result is the highest efficiency reported for any existing OIE-based microfluidic system.In addition,successful co-transfections of three distinct plasmids(EGFP,DsRed and ECFP)into cells were successfully achieved.Hence,we demonstrated that this system is capable of automatically performing multiple gene transfections into mammalian cells.

Thermometry of photosensitive and optically induced electrokinetics chips

Optically induced electrokinetics(OEK)-based technologies,which integrate the high-resolution dynamic addressability of optical tweezers and the high-throughput capability of electrokinetic forces,have been widely used to manipulate,assemble,and separate biological and non-biological entities in parallel on scales ranging from micrometers to nanometers.However,simultaneously introducing optical and electrical energy into an OEK chip may induce a problematic temperature increase,which poses the potential risk of exceeding physiological conditions and thus inducing variations in cell behavior or activity or even irreversible cell damage during bio-manipulation.Here,we systematically measure the temperature distribution and changes in an OEK chip arising from the projected images and applied alternating current(AC)voltage using an infrared camera.We have found that the average temperature of a projected area is influenced by the light color,total illumination area,ratio of lighted regions to the total controlled areas,and amplitude of the AC voltage.As an example,optically induced thermocapillary flow is triggered by the light image-induced temperature gradient on a photosensitive substrate to realize fluidic hydrogel patterning.Our studies show that the projected light pattern needs to be properly designed to satisfy specific application requirements,especially for applications related to cell manipulation and assembly.