Recent advances in dielectrophoresis toward biomarker detection: A summary of studies published between 2014 and 2021.

Dielectrophoresis is a well-understood phenomenon that has been widely utilized in biomedical applications. Recent advancements in miniaturization have contributed to the development of dielectrophoretic-based devices for a wide variety of biomedical applications. In particular, the integration of dielectrophoresis with microfluidics, fluorescence, and electrical impedance has produced devices and techniques that are attractive for screening and diagnosing diseases. This review article summarizes the recent utility of dielectrophoresis in assays of biomarker detection. Common screening and diagnostic biomarkers, such as cellular, protein, and nucleic acid, are discussed. Finally, the potential use of recent developments in machine learning approaches toward improving biomarker detection performance is discussed. This review article will be useful for researchers interested in the recent utility of dielectrophoresis in the detection of biomarkers and for those developing new devices to address current gaps in dielectrophoretic biomarker detection.

Integrated dielectrophoretic and impedimetric biosensor provides a template for universal biomarker sensing in clinical samples.

The detection and quantification of nucleic acid and proteomic biomarkers in bodily fluids is a critical part of many medical screening and diagnoses. However, majority of the current detection platforms are not ideal for routine, rapid, and low-cost testing in point-of-care settings. To address this issue, we developed a concept for a disposable universal point-of-care biosensor that can detect and quantify nucleic acid and proteomic biomarkers in diluted serum samples. The central tenet of sensing is the use of dielectrophoresis, electrothermal effects, and thermophoresis to selectively and rapidly isolate the biomarkers of interest in electrodes and then quantify using electrical impedance. When the sensor was applied to quantify microRNA and antigen biomarker molecules directly in diluted serum samples, it produced a LOD values in the fM range and sensitivity values from 1012 to 1015 Ω/M with a 30 min assay time and assay cost of less than $50 per assay.

Nucleotide Identification in DNA Using Dielectrophoresis Spectroscopy.

We show that negative dielectrophoresis (DEP) spectroscopy is an effective transduction mechanism of a biosensor for the detection of single nucleotide polymorphism (SNP) in a short DNA strand. We observed a frequency dependence of the negative DEP force applied by interdigitated electrodes to polystyrene microspheres (PM) with respect to changes in both the last and the second-to-last nucleotides of a single-strand DNA bound to the PM. The drift velocity of PM functionalized to single-strand DNA, which is proportional to the DEP force, was measured at the frequency range from 0.5 MHz to 2 MHz. The drift velocity was calculated using a custom-made automated software using real time image processing technique. This technology for SNP genotyping has the potential to be used in the diagnosis and the identification of genetic variants associated with diseases.

Integrated dielectrophoretic and surface plasmonic platform for million-fold improvement in the detection of fluorescent events.

We present an integrated dielectrophoretic (DEP) and surface plasmonic technique to quantify ∼1 pM of fluorescent molecules in low conductivity buffers. We have established a DEP force on target molecules to bring those molecules and place them on the nanometallic structures (hotspots) for quantification through surface plasmonic effects. Our results show that the DEP is capable of placing the fluorescent molecules on the hotspots, which are depicted as a significant reduction in the fluorescence lifetime of those molecules. To efficiently integrate the DEP and plasmonic effects, we have designed and utilized pearl-shaped interdigitated electrodes (PIDEs) in experiments. These electrodes generate 2-3 times higher DEP force than traditional interdigitated electrodes. Therefore, high-throughput assays can be developed. The nanometallic structures were strategically fabricated in the periphery of PIDEs for smooth integration of DEP and plasmonic detection. With the introduction of DEP, about 106-fold improvement was achieved over existing plasmonic-based detection. Therefore, this simple addition to the existing surface plasmonic-based detection will enable the disease related protein detection.

Negative dielectrophoresis spectroscopy for rare analyte quantification in biological samples.

We propose the use of negative dielectrophoresis (DEP) spectroscopy as a technique to improve the detection limit of rare analytes in biological samples. We observe a significant dependence of the negative DEP force on functionalized polystyrene beads at the edges of interdigitated electrodes with respect to the frequency of the electric field. We measured this velocity of repulsion for 0% and 0.8% conjugation of avidin with biotin functionalized polystyrene beads with our automated software through real-time image processing that monitors the Rayleigh scattering from the beads. A significant difference in the velocity of the beads was observed in the presence of as little as 80 molecules of avidin per biotin functionalized bead. This technology can be applied in the detection and quantification of rare analytes that can be useful in the diagnosis and the treatment of diseases, such as cancer and myocardial infarction, with the use of polystyrene beads functionalized with antibodies for the target biomarkers.

Dielectrophoretic label-free immunoassay for rare-analyte quantification in biological samples.

The current gold standard for detecting or quantifying target analytes from blood samples is the ELISA (enzyme-linked immunosorbent assay). The detection limit of ELISA is about 250 pg/ml. However, to quantify analytes that are related to various stages of tumors including early detection requires detecting well below the current limit of the ELISA test. For example, Interleukin 6 (IL-6) levels of early oral cancer patients are <100 pg/ml and the prostate specific antigen level of the early stage of prostate cancer is about 1 ng/ml. Further, it has been reported that there are significantly less than 1pg/mL of analytes in the early stage of tumors. Therefore, depending on the tumor type and the stage of the tumors, it is required to quantify various levels of analytes ranging from ng/ml to pg/ml. To accommodate these critical needs in the current diagnosis, there is a need for a technique that has a large dynamic range with an ability to detect extremely low levels of target analytes (