Devices and methods for increasing throughput of flow-based electroporation systems

A method of electroporation using a microfluidic device, the method comprising:

• flowing a liquid through a channel in the microfluidic device in a flow direction, the fluid including a plurality of cells and molecules, wherein the channel comprises a channel height (H) and a channel width (W), the ratio of the channel height to the channel width being less than about 0.1;
• generating a time-dependent electric field across the channel and perpendicular to the flow direction using electrodes arranged transverse to the direction of flow, wherein a polarity of the time-dependent electric field is configured to alternate as a function of time, and wherein a waveform of the time-dependent electric field comprises a series of alternating positive and negative peaks;
• and
• coordinating a flow speed of the liquid through the channel and a waveform of the time-dependent voltage applied to the electrodes to generate the time-dependent electric field such that the molecules are moved into the plurality of cells after being subject to the time-dependent electric field, wherein the voltage wave form comprises a periodic bipolar waveform with alternating segments of symmetric reversed polarity, each of the alternating segments having a first segment portion followed by a second segment portion, the first segment portion having a shorter duration and a higher amplitude relative to the second segment portion.