ABSTRACT Single cylindrical submicron pores in PMMA polymer membranes are micropatterned by electron beam lithography and integrated into all PMMA-based electrophoretic flow detector systems. Pore dimensions are 450 nm in diameter and 1 μm in length. The pores are electrically characterized in aqueous KCl electrolyte, exhibiting a stable time-independent ionic current through the pore with a noise level of less than 1% of the open-pore current. The current-voltage curves are linear and scale with electrolyte concentration. The negative surface charge of the membrane over-proportionally decreases pore conductance at low electrolyte concentrations (≤0.1 M) that are still beyond those typically applied in biological experiments. Pores do not exhibit rectification of current flowing through them, allowing for operation with either polarity. To allow for detection of yet much smaller particles, the described PMMA-based system also was successfully equipped with pores of 1.5 nm instead of 450 nm in diameter. This was achieved by introducing naturally occurring biological protein pores of α-hemolysin on a lipid bilayer into the prepatterned PMMA membrane of an assembled PMMA-based electrophoretic flow detector system. Characteristics of translocation events of single-stranded linear plasmid DNA molecules through the pores were recorded, and ionic current deductions during biomolecule translocation were clear and distinguished. Based on the presented submicron scale open pore ionic current transport properties, as well as the observed passage of DNA molecules through protein pores inserted into PMMA membranes, our current research proposes that all PMMA electrophoretic flow detectors exhibit an excellent potential for future use as biomedical resistive-pulse sensors, as long as pore dimensions match those of biomolecules to be detected.
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