Optimization of Electrolytes with Redox Reagents to Improve the Impedimetric Signal for Use with a Low-Cost Analyzer

The most well-known criterion for POC devices is ASSURED, and affordability, i.e., using low-cost instrumentation, is the most challenging one. This manuscript provides a pathway for transitioning ESSENCE, an impedance-based biosensor platform, from using an expensive benchtop analyzer—KeySight 4294A (~$50k)—to using a significantly portable and cheaper USB oscilloscope—Analog Discovery 2 (~$200) —with similar sensitivity (around 100 times price difference). To achieve this, we carried out a fundamental study of the interplay between an electrolyte like potassium chloride (KCl), and an electrolyte buffer like phosphate buffered saline (PBS) in the presence and absence of a redox buffer like ferro/ferricyanide system and ([Ru(bpy)₃]²⁺). Redox molecules in the electrolyte caused a significant change in the Nyquist curve of the impedance depending on the redox molecule type. The redox species and the background electrolyte have their own RC semicircles in the Nyquist curve, whose overlap depends on the redox concentration and electrolyte ionic strength. We found that by increasing the electrolyte ionic strength or the redox concentration, the RC semicircle moves to higher frequencies and vice versa. Importantly, the use of the buffer electrolyte, instead of KCl, led to a lower standard deviation and overall signal (lesser sensitivity). However, to achieve the best results from the biorecognition signal, we chose a buffered electrolyte like PBS with high ionic strength and lowered the redox probe concentrations to minimize the standard deviation and reduce any noise from migrating to the low-cost analyzer. Comparing the two analyzers shows similar results, with a lowered detection limit from the low-cost analyzer.