AC dielectrophoretic manipulation and electroporation of vaccinia virus using carbon nanoelectrode arrays

Abstract

This paper reports the capture and detection of vaccinia virus particles based on AC dielectrophoresis (DEP) and electrochemical impedance measurements employing an embedded vertically aligned carbon nanofiber (VACNF) nanoelectrode array (NEA) versus a macroscopic indium-tin-oxide (ITO) transparent electrode in a "points-and-lid" configuration. The nano-DEP device was fabricated by bonding two SU-8 covered electrodes patterned using photolithography. The bottom electrode contains a 200 × 200 μm active region on a randomly distributed NEA and the top electrode contains a microfluidic channel in SU-8 spin-coated on ITO to guide the flow of the virus solution. The real-time impedance change was measured during DEP capture and validated with fluorescence microscopy measurements. The NEA was able to capture virus particles with a rather low AC voltage (∼8.0 V peak-to-peak) at 1.0 kHz frequency as the particles were passed through the fluidic channel at high flow velocities (up to 8.0 mm/s). A concentration detection limit as low as ∼2.58 × 10 particles/mL was obtained via impedance measurements after only 54 sec of DEP capture. At the low AC frequencies (50.0 Hz or less), the high electric field at the exposed VACNF tips induced electroporation of the DEP-captured virus particles, which was validated by fluorescence emission from the dyes staining lipophilic membrane and internal nucleic acid, respectively. This study suggests the possibility of integration of a fully functional electronic device for rapid, reversible and label-free capture and detection of pathogenic viruses, with a potential of generating electroporation to the captured the virus particles for further biochemical study.

Analysis of microorganisms with nonlinear electrokinetic microsystems

“…Going even further, the development of nontraditional materials has allowed scientists to examine viruses in a new way, as is seen in the work of Madiyar et al. [40]. The basis of manipulation is with AC DEP, but the arrays are made of carbon nanofibers creating a nanoelectrode array [40].…”

“…[40]. The basis of manipulation is with AC DEP, but the arrays are made of carbon nanofibers creating a nanoelectrode array [40]. The design and material allowed for the capture of viral particles at a lower AC voltage of 8.0 V with a 1.0 kHz frequency; additionally, the group reported that at 50 Hz or less, electroporation can occur due to the electric field at the carbon nanofiber tips [40].…”

Dielectrophoresis applications in biomedical field and future perspectives in biomedical technology

“…The application of DEP to the biomedical field has recently gained significant attention. DEP can be used in several areas including cancer disease diagnostics [36], stem cell‐based therapy [32,42], and diagnosis of infectious diseases [30,43,44]. In this review, the recent applications of DEP to the biomedical field are discussed.…”

Analysis of Bacteriophages with Insulator-Based Dielectrophoresis

“…Singh et al created a sensor for influenza virus employing carbon nanotubes that were electrodeposited by means of DEP [27]. Madiyar et al reported the capture and detection of vaccinia virus with DEP by using carbon nanoelectrode arrays [28]. Some earlier studies involved plant viruses.…”