A Solid‐State Hard Microfluidic–Nanopore Biosensor with Multilayer Fluidics and On‐Chip Bioassay/Purification Chamber

Abstract

Solid-state nanopores are an emerging biosensor for nucleic acid and protein characterization. For use in a clinical setting, solid-state nanopore sensing requires sample preparation and purification, fluid handling, a heating element, electrical noise insulators, and an electrical readout detector, all of which hamper its translation to a point-of-care diagnostic device. A stand-alone microfluidicbased nanopore device is described that combines a bioassay reaction/purification chamber with a solid-state nanopore sensor. The microfluidic device is composed of the high-temperature/solvent resistance Zeonex plastic, formed via micro-machining and heat bonding, enabling the use of both a heat regulator and a magnetic controller. Fluid control through the microfluidic channels and chambers is controlled via fluid port selector valves and allows up-to eight different solutions. Electrical noise measurements and DNA translocation experiments demonstrate the integrity of the device, with performance comparable to a conventional stand-alone nanopore setup. However, the microfluidic-nanopore setup is superior in terms of ease of use. To showcase the utility of the device, single molecule detection of a DNA PCR product, after magnetic bead DNA separation, is accomplished on chip.

Digital immunoassay for biomarker concentration quantification using solid-state nanopores

“…Likewise, the sensitivity of this assay, currently limited by the lower bound of the concentration ratio of junction strand-toprobe, x ¼ c Js c P % 0:01; can be adjusted to accommodate any desired concentration range at the cost of linearly increasing the detection time of a single nanopore sensor as concentrations are reduced, though this increase in counting time can be offset through parallelization with an array of pores and other strategies that increase nanopore capture rate 62 . The limit of detection, while fixed by the parameters of a particular assay and the choice of probe concentration, can therefore be controlled by the counting time of the nanopore and for a fixed measurement time can be improved by speeding up the detection through parallelization, amplification, preconcentration, or capture rate enhancement schemes 19,29,53,[62][63][64][65][66][67][68] .…”

A click chemistry amplified nanopore assay for ultrasensitive quantification of HIV-1 p24 antigen in clinical samples

“…In addition, the current assay protocol still involves multiple hands-on steps, leading to long turn-around and potential human errors. Assay automation by microfluidic technologies needs to be incorporated to reduce both time and financial costs of this method, especially for future point-ofcare applications 81 . By addressing these issues, we believe that the CAN assay can afford an ultrasensitive, rapid and robust method for HIV-1 early diagnosis and prognosis, and can also be applied to mitigate any future infectious disease outbreaks with minimum modification.…”

A Critical Review on the Sensing, Control, and Manipulation of Single Molecules on Optofluidic Devices

“…A number of efforts are reported to develop on-chip lasers sources [437][438][439][440] that can ease the optical integration and potentially eliminate the complex optical alignment required to carry out the SM studies. Significant advances have been made toward on-chip sample preparation [271,[441][442][443] for developing the next-generation point-of-care (POC) detection system [444][445][446][447][448]. Hybrid optofluidic integration that combines PDMS and silicon-based optofluidic devices is also opening avenues and allowing more flexibility, reconfigurability, and adaptability for SM studies [441].…”