Single-neuronal cell culture and monitoring platform using a fully transparent microfluidic DEP device

Abstract

Dielectrophoresis using multi-electrode arrays allows a non-invasive interface with biological cells for long-term monitoring of electrophysiological parameters as well as a label-free and non-destructive technique for neuronal cell manipulation. However, experiments for neuronal cell manipulation utilizing dielectrophoresis have been constrained because dielectrophoresis devices generally function outside of the controlled environment (i.e. incubator) during the cell manipulation process, which is problematic because neurons are highly susceptible to the properties of the physiochemical environment. Furthermore, the conventional multi-electrode arrays designed to generate dielectrophoretic force are often fabricated with non-transparent materials that confound live-cell imaging. Here we present an advanced single-neuronal cell culture and monitoring platform using a fully transparent microfluidic dielectrophoresis device for the unabated monitoring of neuronal cell development and function. The device is mounted inside a sealed incubation chamber to ensure improved homeostatic conditions and reduced contamination risk. Consequently, we successfully trap and culture single neurons on a desired location and monitor their growth process over a week. The proposed single-neuronal cell culture and monitoring platform not only has significant potential to realize an in vitro ordered neuronal network, but also offers a useful tool for a wide range of neurological research and electrophysiological studies of neuronal networks.

A Single-Neuron: Current Trends and Future Prospects

“…The DEP electrode array traps and releases neurons (one at a time/electrode), as shown in Figure 20. The segregated single-neurons can be cultured and monitored over time, allowing the screening of various electrophysiological parameters and enabling detailed neurological studies [58].…”

“…The trapped neuron was recorded, and its morphological changes were tracked with the assist of a phasecontrast microscope. Thus, this device enabled us to study multiple single-neurons at the same time and also electrical communication between them [58]. The first neurochip was a silicon-based micromachined device with a 4 × 4 array of metal electrodes, which allowed growth and monitor neuronal cell individually.…”

“…The trapped neuron was recorded, and its morphological changes were tracked with the assist of a phase-contrast microscope. Thus, this device enabled us to study multiple single-neurons at the same time and also electrical communication between them [58].…”

“…At the end, when the incoming neuron reaches the outside of the electrode, the repulsive force pushed the neuron out of the ring. Reprinted with permission from[58].…”

Compartmentalized Neuronal Culture for Viral Transport Research

“…Application of microfluidics in the field of cell biology is becoming a powerful tool, due to its ability to precisely control, manipulate, and monitor sub-millimeter volume cellular culture environments (Taylor et al, 2005;Whitesides, 2006). Initially, microfluidic devices were designed for chemical and biomolecular analyses with higher sensitivity than traditional methods (Atencia and Beebe, 2005;Kim et al, 2018); more recently, they have been applied in cell biology (Dexter and Parker, 2009;Lo and Yao, 2015;Zhao et al, 2019).…”