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Mechanism and kinetics of lipid bilayer formation in solid-state nanopores
Uppsala University, Disciplinary Domain of Science and Technology, Technology, Department of Engineering Sciences, Solid State Electronics. Uppsala University.
Uppsala University, Disciplinary Domain of Science and Technology, Technology, Department of Engineering Sciences, Solid State Electronics.ORCID iD: 0000-0003-4948-8353
Linköping University.
Uppsala University, Disciplinary Domain of Science and Technology, Technology, Department of Engineering Sciences, Solid State Electronics.ORCID iD: 0000-0003-4395-7905
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2020 (English)In: Langmuir, ISSN 0743-7463, E-ISSN 1520-5827Article in journal (Refereed) Published
Abstract [en]

Solid-state nanopores provide a highly versatile platform for rapid electrical detection and analysis of single molecules. Lipid bilayer coating of the nanopores can reduce non-specific analyte adsorption to the nanopore sidewalls and increase the sensing selectivity by providing possibilities for tethering specific ligands in a cell-membrane mimicking environment. However, mechanism and kinetics of lipid bilayer formation from vesicles remain unclear in the presence of nanopores. In this work, we used a silicon-based, truncated-pyramidal nanopore array as the support for lipid bilayer formation. Lipid bilayer formation in the nanopores was monitored in real-time by the change in ionic current through the nanopores. Statistical analysis revealed that a lipid bilayer is formed from instantaneous rupture of individual vesicle upon adsorption in the nanopores, differing from the generally agreed mechanism that lipid bilayers form at a high vesicle surface coverage on a planar support. The2dependence of the lipid-bilayer formation process as a function of applied bias, vesicle size and concentration was systematically studied. In addition, the non-fouling properties of the lipid bilayer coated nanopores were demonstrated during long single stranded DNA translocation through the nanopore array. The findings indicate that lipid bilayer formation process can be modulated by introducing nanocavities intentionally on the planar surface to create active sites or changing the vesicle size and concentration.

Place, publisher, year, edition, pages
2020.
National Category
Engineering and Technology
Identifiers
URN: urn:nbn:se:uu:diva-399725DOI: 10.1021/acs.langmuir.9b03637OAI: oai:DiVA.org:uu-399725DiVA, id: diva2:1379057
Available from: 2019-12-16 Created: 2019-12-16 Last updated: 2020-02-06Bibliographically approved
In thesis
1. Solid-state nanopores: fabrication and applications
Open this publication in new window or tab >>Solid-state nanopores: fabrication and applications
2020 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Nanopores are of great interest in study of DNA sequencing, protein profiling and power generation. Among them, solid-state nanopores show obvious advantages over their biological counterparts in terms of high chemical stability and reusability as well as compatibility with the existing CMOS fabrication techniques. Nanopore sensing is most frequently based on measuring ionic current through a nanopore while applying a voltage across it. When an analyte passes through the pore, the ionic current temporarily changes, providing information of the analyte such as its size, shape and surface charge. Although many magnificent reports on using solid-state nanopores have appeared in the literature, several challenges still remain for their wider applications, which include improvement of fabrication reproducibility for mass production of ultra-small nanopores and minimization of measurement instability as well as control of translocation speed and reduction of background noise. This thesis work explores different techniques to achieve robust and high throughput fabrication of sub-10 nm nanopores for different applications.

The thesis starts with presenting various fabrication techniques explored during my PhD studies. Focused ion beam method was firstly employed to drill nanopores in free-standing SiNx membranes. Sub-10 nm nanopores could be obtained with a focused helium ion beam. But the fabrication throughput was limited with this technique. A new fabrication process combing electron beam lithography (EBL) with reactive ion etching/ion beam etching, which is compatible with the existing CMOS fabrication technology, was developed to realize a high throughput, mass production of nanopores in free-standing SiNx membranes. However, the smallest size that could be controllably achieved with this process was around 40 nm, which is still far from sub-10 nm in size required for, e.g., DNA sequencing. Finally, by using anisotropic etching of single-crystal silicon in KOH solution, sub-5 nm truncated pyramidal nanopores were mass produced with good process controllability in a silicon-on-insulator (SOI) substrate. In addition, nanopore arrays were also successfully fabricated using a modified EBL based fabrication process.

Then, several sensing application examples using either single nanopores or nanopore arrays were investigated. Translocation of nanoparticles, DNA and proteins were demonstrated using the fabricated single nanopores or nanopore arrays in a single freestanding membrane. Moreover, the kinetics and mechanism of the lipid bilayer formation in nanopore array, aiming to prevent non-specific adsorption, were studied using ionic current measurements. In addition, individual addressability of a solid-state nanopore array on separated freestanding membranes was realized by integrating microfluidics and a customized multiplexer.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2020. p. 82
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 1890
Keywords
solid-state nanopore, truncated-pyramidal nanopore, nanopore array, pore size reduction, individual addressability, microfluidics, translocation.
National Category
Engineering and Technology
Research subject
Engineering Science with specialization in Electronics
Identifiers
urn:nbn:se:uu:diva-399726 (URN)978-91-513-0838-8 (ISBN)
Public defence
2020-02-21, Polhemsalen, Ångströmlaboratoriet, Lägerhyddsvägen 1, Uppsala, 09:15 (English)
Opponent
Supervisors
Available from: 2020-01-31 Created: 2019-12-16 Last updated: 2020-01-31

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Zeng, ShuangshuangLi, ShiyuWen, ChenyuZhang, Shi-LiZhang, Zhen

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