Pressure-Biased Nanopores for Excluded Volume Metrology, Lipid Biomechanics, and Cell-Adhesion Rupturing

Nanopore sensing has been widely used in applications ranging from DNA sequencing to disease diagnosis. To improve these capabilities, pressure-biased nanopores have been explored in the past to-primarily-increase the residence time of the analyte inside the pore. Here, we studied the effect of pressure on the ability to accurately quantify the excluded volume which depends on the current drop magnitude produced by a single entity. Using the calibration standard, the inverse current drop (1/ΔI) decreases linearly with increasing pressure, while the dwell drop reduces exponentially. We therefore had to derive a pressure-corrected excluded volume equation to accurately assess the volume of translocating species under applied pressure. Moreover, a method to probe deformation in nanoliposomes and a single cell is developed as a result. We show that the soft nanoliposomes and even cells deform significantly under applied pressure which can be probed in terms of the shape factor which was introduced in the excluded volume equation. The proposed work has practical applications in mechanobiology, namely, assessing the stiffness and mechanical rigidity of liposomal drug carriers. Pressure-biased pores also enabled multiple observations of cell-cell aggregates as well as their subsequent rupture, potentially allowing for the study of microbial symbioses or pathogen recognition by the human immune system.