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.
Dielectrophoresis (DEP) is a commonly used technique in biomedical engineering to manipulate biomolecules. DEP is defined as the force acting on dielectric particles when they are exposed to non-uniform electric fields. DEP effect can be divided in three categories: positive (dielectric particles are attracted to the electrodes), negative, and zero force DEP. The cross-over frequency is the frequency in which the DEP force is equal to zero. The cross-over frequency depends on the conductivity and the permittivity of the particles and of the suspended medium. The DEP cross-over frequency has been utilized in detecting/quantifying biomolecules. A manual procedure is commonly used to estimate the cross-over frequency of biomolecules. Therefore, the accuracy of this detection method is significantly limited. To address this issue, we designed and tested an automated procedure to carry out DEP spectroscopy in dielectric particles dissolved in a biological buffer solution. Our method efficiently measures the effect of the DEP force through a live video feed from the microscope camera and performs real-time image processing. It records the change in the fluorescence emission as the system automatically scans the electric frequency of the function generator over a specified time interval. We demonstrated the effectiveness of the method by extracting the crossover frequencies and the DEP spectrum of polystyrene beads with blue color dye (1000 nm diameter) and green fluorescent polystyrene beads with 500 nm diameter using this procedure. This approach can lead to the development of a biosensor with significantly higher sensitivity than existing detection methods.