Passivated-electrode insulator-based dielectrophoretic separation of heterogeneous cell mixtures.

Rapid and accurate purification of various heterogeneous mixtures is a critical step for a multitude of molecular, chemical, and biological applications. Dielectrophoresis has shown to be a promising technique for particle separation due to its exploitation of the intrinsic electrical properties, simple fabrication, and low cost. Here, we present a geometrically novel dielectrophoretic channel design which utilizes an array of localized electric fields to separate a variety of unique particle mixtures into distinct populations. This label-free device incorporates multiple winding rows with several nonuniform structures on to sidewalls to produce high electric field gradients, enabling high locally generated dielectrophoretic forces. A balance between dielectrophoretic forces and Stokes' drag is used to effectively isolate each particle population. Mixtures of polystyrene beads (500 nm and 2 µm), breast cancer cells spiked in whole blood, and for the first time, neuron and satellite glial cells were used to study the separation capabilities of the design. We found that our device was able to rapidly separate unique particle populations with over 90% separation yields for each investigated mixture. The unique architecture of the device uses passivated-electrode insulator-based dielectrophoresis in an innovative microfluidic device to separate a variety of heterogeneous mixture without particle saturation in the channel.

Breast cancer cell obatoclax response characterization using passivated-electrode insulator-based dielectrophoresis.

Inherent electrical properties of cells can be beneficial to characterize different cell lines and their response to experimental drugs. This paper presents a novel method to characterize the response of breast cancer cells to drug stimuli through use of off-chip passivated-electrode insulator-based dielectrophoresis (OπDEP) and the application of AC electric fields. This work is the first to demonstrate the ability of OπDEP to differentiate between two closely related breast cancer cell lines, LCC1 and LCC9 while assessing their drug sensitivity to an experimental anti-cancer agent, Obatoclax. Although both cell lines are derivatives of estrogen-responsive MCF-7 breast cancer cells, growth of LCC1 is estrogen independent and anti-estrogen responsive, while LCC9 is both estrogen-independent and anti-estrogen resistant. Under the same operating conditions, LCC1 and LCC9 had different DEP profiles. LCC1 cells had a trapping onset (crossover) frequency of 700 kHz and trapping efficiencies between 30-40%, while LCC9 cells had a lower crossover frequency (100 kHz) and showed higher trapping efficiencies of 40-60%. When exposed to the Obatoclax, both cell lines exhibited dose-dependent shifts in DEP crossover frequency and trapping efficiency. Here, DEP results supplemented with cell morphology and proliferation assays help us to understand the response of these breast cancer cells to Obatoclax.

Rapid, low-cost fabrication of electronic microfluidics via inkjet-printing and xurography (MINX).

Microfluidics has shown great promise for point-of-care assays due to unique chemical and physical advantages that occur at the micron scale. Furthermore, integration of electrodes into microfluidic systems provides additional capabilities for assay operation and electronic readout. However, while these systems are abundant in biological and biomedical research settings, translation of microfluidic devices with embedded electrodes are limited. In part, this is due to the reliance on expensive, inaccessible, and laborious microfabrication techniques. Although innovative prior work has simplified microfluidic fabrication or inexpensively patterned electrodes, low-cost, accessible, and robust methods to incorporate all these elements are lacking. Here, we present MINX, a low-cost <1 USD and rapid (∼minutes) fabrication technique to manufacture microfluidic device with embedded electrodes. We characterize the structures created using MINX, and then demonstrate the utility of the approach by using MINX to implement an electrochemical bead-based biomarker detection assay. We show that the MINX technique enables the scalable, inexpensive fabrication of microfluidic devices with electronic sensors using widely accessible desktop machines and low-cost materials.

A sample-to-answer electrochemical biosensor system for biomarker detection.

Biomarker detection is critical for the diagnosis and treatment of numerous diseases. Typically, target biomarkers in blood samples are measured through tests conducted at centralized laboratories. Testing at central laboratories increases wait times for results, in turn increasing healthcare costs and negatively impacting patient outcomes. Alternatively, point-of-care platforms enable the rapid measurement of biomarkers, expand testing location capabilities and mitigate manual processing steps through integration and automation. However, many of these systems focus on sample detection rather than the equally important sample preparation. Here we present a fully integrated and automated sample-to-answer electrochemical biosensing platform which incorporates each aspect of the biomarker testing workflow from blood collection to sample preparation to assay operation and readout. The system combines a commercial microneedle blood sampling device with membrane-based plasma filtration upstream of a bead-based electrochemical immunoassay. We characterize the high separation efficiency (>99%) and low non-specific binding of the whole blood-to-plasma filtration membrane under a range of operating conditions. We demonstrate a full sample-to-answer workflow through the analysis of interlukin-6-spiked blood samples.

Deactivation of E. coli in water using Fe3+-saturated montmorillonite impregnated filter paper.

In areas with high exposure to pathogen contaminated water and lack the economic means for water treatment, low cost and convenient point-of-use drinking water disinfection materials/devices are essential. Using a simple craft paper making method, Fe3+-saturated montmorillonite impregnated filter paper was constructed to filter live Escherichia coli (E. coli)-spiked water. The Scanning Electron Microscopic images of the E. coli cells in contact with the Fe3+-saturated montmorillonite impregnated filter paper showed: 1) Fe3+-saturated montmorillonite particles were uniformly coated on the cellulose paper fiber, creating large mineral surface for cell contact; and 2) E. coli cell membrane was dehydrated and damaged, resulting cell deactivation upon contacting with the Fe3+-saturated montmorillonite particles impregnated in the paper. The E. coli cells passing through the Fe3+-saturated montmorillonite impregnated filter paper were not viable as further confirmed by the microfluidic dielectrophoresis analysis. They remained non-viable at room temperature even after 5 days, as shown by the results from both the Colony Counting test and the Colilert test. More than 99.5% deactivation efficiency was achieved when the ratio of the volume of the E. coli contaminated water to the mass of Fe3+-saturated montmorillonite was maintained at <1:1.5 (mL/mg). The Fe3+-saturated montmorillonite impregnated filter paper maintained ~74% E. coli deactivation efficiency even after the 8th consecutive use. About 0.52 mg Fe3+, which is bioavailable, could be leached into the water for every 2 L E coli-contaminated water that is treated with the filter paper. The treated water could therefore provide iron supplement to a person at a level within the range of the FDA recommended human daily intake of iron. The results from this study has clearly demonstrated promising potential of using the Fe3+-saturated montmorillonite impregnated filter paper for low cost (~$0.07/L treated water for this study) and convenient point-of-use drinking water disinfection.