The paper presents a novel microfluidic device for identification and characterization of cells in suspensions using impedance spectroscopy. The device consists of two glass wafers: a bottom wafer comprising a microfluidic channel with two electrodes added for impedance measurement, and a top glass wafer in which inlets and outlets are realized. The fact that the device is glass-based provides a few key advantages: reduced influence from parasitic components during measurements (due to the good isolation properties of the substrate), optical transparency and hydrophilic surface of the microfluidic channel. The latter feature is especially important as it enables sample suction due to capillarity forces only. Thus, no external pumping is required and only a small volume sample suffices for the measurement.
The fabrication process of this device consists of three major steps. First, via-holes and inlet/outlet holes are executed in the top glass wafer by wet etching in a 49% HF solution using a low stress amorphous silicon/silicon carbide/photoresist mask. Second, the microfluidic channel is etched into the bottom wafer and Ti/Pt electrodes are then patterned on top of it using a spray coating-based lithography. The last processing step is bonding together the top and bottom glass wafers by employing a very thin adhesive intermediate layer (SU8). This adhesive layer was applied selectively only on the bottom die, from a Teflon cylinder, using a contact imprinting method.
Finally, fabricated devices were successfully tested using DI water, phosphate buffer saline (PBS), and various types of both dead cells and living cells resuspended in PBS. Clear differences between dead and live cells have been observed. The impedance measurements were carried out in the frequency range 5 kHz to 10 MHz. The measured magnitude and phase were studied using different types of cells in Dulbecco's Minimal Essential medium (DMEM). The obtained impedance spectra revealed the characteristic spectra signature for each type of cell.