A low voltage, two-level-metal, and multi-layer insulator electrowetting-on-dielectric (EWD) platform is presented. Dispensing 300pl droplets from 140nl closed on-chip reservoirs was accomplished with as little as 11.4V solely through EWD forces, and the actuation threshold voltage was 7.2V with a 1Hz voltage switching rate between electrodes. EWD devices were fabricated with a multilayer insulator consisting of 135nm sputtered tantalum pentoxide (Ta 2 O 5 ) and 180nm parylene C coated with 70nm of CYTOP. Furthermore, the minimum actuation threshold voltage followed a previously published scaling model for the threshold voltage, V T , which is proportional to (t/ε r ) 1/2 , where t and ε r are the insulator thickness and dielectric constant respectively. Device threshold voltages are compared for several insulator thicknesses (200nm, 500nm, and 1µm), different dielectric materials (parylene C and tantalum pentoxide), and homogeneous versus heterogeneous compositions. Additionally, we used a two-level-metal fabrication process, which enables the fabrication of smaller and denser electrodes with high interconnect routing flexibility. We also have achieved low dispensing and actuation voltages for scaled devices with 30pl droplets.
Picoliter droplets actuated on an electrowetting-on-dielectric (EWD) actuator are demonstrated. In this study, the physical scaling of electrodes for 33 μm and 21 μm EWD devices resulted in droplets of 12 pl and 5 pl being dispensed respectively in conjunction with 3 μm SU8 gaskets. The stacked multi-layer insulators in the actuators consisted of 200 nm tantalum pentoxide (Ta2O5) and 200 nm parylene C films deposited and coated with 70 nm of CYTOP. The voltages for dispensing droplets on chips without any external pressure sources are 17.1 Vrms and 22 Vrms for these two sets of devices. A 12 pl droplet can be split into two 6 pl daughter droplets at 18.7 Vrms with 33 μm electrode devices. Droplet manipulation is also demonstrated with paramagnetic beads and buffer solutions with proteins. In addition, electrodes with interlocking protrusions and special featured reservoir gasket are designed to facilitate droplet dispensing on these scaled EWD devices. In order to improve sealing of the two-piece sandwich EWD structure, a soft material, Norland Optical Adhesive (NOA), was coated on the top plate along with pressure on top. We demonstrate that based on fundamental theories and experiments, the dimensional scaling of EWD devices has not yet met a limitation as long as the EWD device can be sealed well.
The results of investigations into performing DNA sequencing chemistry on a picoliter-scale electrowetting digital microfluidic platform are reported. Pyrosequencing utilizes pyrophosphate produced during nucleotide base addition to initiate a process ending with detection through a chemiluminescence reaction using firefly luciferase. The intensity of light produced during the reaction can be quantified to determine the number of bases added to the DNA strand. The logic-based control and discrete fluid droplets of a digital microfluidic device lend themselves well to the pyrosequencing process. Bead-bound DNA is magnetically held in a single location, and wash or reagent droplets added or split from it to circumvent product dilution. Here we discuss the dispensing, control, and magnetic manipulation of the paramagnetic beads used to hold target DNA. We also demonstrate and characterize the picoliter-scale reaction of luciferase with adenosine triphosphate to represent the detection steps of pyrosequencing and all necessary alterations for working on this scale.
The development of a photothermal absorbance detector for use with microfluidic devices is described. Unlike thermooptical techniques that rely on measuring refractive index changes, the solution viscosity is probed by continuously monitoring solution conductivity. Platinum electrodes microfabricated on a quartz substrate and bonded to a substrate containing the microchannels enable contact conductivity measurements. The effects of excitation frequency and voltage, electrode spacing, laser power, and laser modulation (chopping) frequency were evaluated experimentally. In the current configuration a limit of detection of 5 nM for DABSYL-tagged glucosamine was obtained using long injections (to give flat-topped peaks). This corresponds to an absorbance of 4.4 × 10−7 AU. Separation and detection of DABSYL-tagged glycine, proline, and tryptophan is also shown to demonstrate the feasibility of the method. In addition, simulations were used to investigate the applicability of the technique to small volume platforms.
The development and application of polyelectrolytic gel electrodes (PGEs) for a microfluidic photothermal absorbance detection system is described. The PGEs are used to measure changes in conductivity based on heat generation by analytes absorbing light and changing the solution viscosity. The PGEs are suitable for direct contact conductivity measurements since they do not degrade with exposure to high electric fields. Both a 2-electrode system with DC voltages and a 3-electrode system with AC voltages were investigated. Experimental factors including excitation voltage, excitation frequency, laser modulation frequency, laser power, and path length were tested. The limits of detection for the 3-electrode and 2-electrode systems are 500 nM and 0.55 nM for DABSYL-tagged glucosamine, respectively. In addition, an electrokinetic separation of a potassium, DABSYL-tagged glucosamine, Rhodamine 6G, and Rhodamine B mixture was demonstrated.
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