Abstract
Insulator-based dielectrophoresis can be used to manipulate biological particles, but has thus far found limited practical applications due to low sensitivity. We present linear sweep three-dimensional insulator-based dielectrophoresis as a considerably more sensitive approach for strain-level discrimination bacteria. In this work, linear sweep three-dimensional insulator-based dielectrophoresis was performed on Pseudomonas aeruginosa PA14 along with six isogenic mutants as well as Streptococcus mitis SF100 and PS344. Strain-level discrimination was achieved between these clinically important pathogens with applied electric fields below 10 V/mm. This low voltage, high sensitivity technique has potential applications in clinical diagnostics as well as microbial physiology research.
Highlights
The cell envelope properties of bacteria play a central role in cellular function [1]
P. aeruginosa biofilms have been extensively studied in the context of cystic fibrosis, where biofilm formation can play a key role in patient morbidity and mortality [3,4,5,6,7,8,9], as well fluid mechanics, with recent work demonstrating that particular hydrodynamic conditions can stimulate the formation of biofilm streamers [10]
Aggregation Experiments at Constant Voltage We found that strains with a proclivity for biofilm formation assemble into tightly bound aggregates during 3DiDEP immobilization
Summary
The cell envelope properties of bacteria play a central role in cellular function [1]. This phenomena has previously been used to induce the formation of bacterial aggregates [16], but previous investigations have been hampered by low sensitivity and high applied voltages, resulting in damage to the cells and only coarse discrimination between cells with very different surface properties such as Gram-positive versus Gram-negative cells [15]. We demonstrate linear sweep 3DiDEP based phenotyping to measure physiological properties associated with virulence factors and biofilm formation in P. aeruginosa and S. mitis.
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