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Teflon-on-Glass Molding Enables High-Throughput Fabrication of Hydrophilic-in-Hydrophobic Microwells for Bead-Based Digital Bioassays

In recent years, Teflon-on-glass microwells have been successfully implemented in bead-based digital bioassays for the sensitive detection of single target molecules. Their hydrophilic-in-hydrophobic (HIH) nature enables the isolation and analysis of individual beads, carrying the target molecules,...

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Autores principales: Tripodi, Lisa, Ven, Karen, Kil, Dries, Rutten, Iene, Puers, Robert, Lammertyn, Jeroen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6266592/
https://www.ncbi.nlm.nih.gov/pubmed/30388800
http://dx.doi.org/10.3390/ma11112154
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author Tripodi, Lisa
Ven, Karen
Kil, Dries
Rutten, Iene
Puers, Robert
Lammertyn, Jeroen
author_facet Tripodi, Lisa
Ven, Karen
Kil, Dries
Rutten, Iene
Puers, Robert
Lammertyn, Jeroen
author_sort Tripodi, Lisa
collection PubMed
description In recent years, Teflon-on-glass microwells have been successfully implemented in bead-based digital bioassays for the sensitive detection of single target molecules. Their hydrophilic-in-hydrophobic (HIH) nature enables the isolation and analysis of individual beads, carrying the target molecules, which can be further manipulated accurately through optical tweezer (OT) setups. However, these Teflon HIH-microwell platforms are conventionally fabricated through a complex, time-consuming and labor-intensive dry lift-off procedure which involves a series of major steps, limiting the up-scaling potential of these platforms. Alternative Teflon-based microwell fabrication methods have been extensively explored in literature but they preclude the generation of hydrophobic wells with hydrophilic bottom, thereby hampering the bioassay performance. Here, we present a new Teflon-on-glass molding method for the high throughput fabrication of hydrophilic-in-hydrophobic (HIH) microwell arrays, able to empower bead-based digital bioassays. Microwells 2.95 μm in depth and 3.86 μm in diameter were obtained to host individual beads. In these microwell arrays, sealing of reagents was demonstrated with an efficiency of 100% and seeding of superparamagnetic beads was achieved with an efficiency of 99.6%. The proposed method requires half as many steps when compared to the traditional dry lift-off process, is freely scalable and has the potential to be implemented in different bead-based bioassay applications.
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spelling pubmed-62665922018-12-17 Teflon-on-Glass Molding Enables High-Throughput Fabrication of Hydrophilic-in-Hydrophobic Microwells for Bead-Based Digital Bioassays Tripodi, Lisa Ven, Karen Kil, Dries Rutten, Iene Puers, Robert Lammertyn, Jeroen Materials (Basel) Article In recent years, Teflon-on-glass microwells have been successfully implemented in bead-based digital bioassays for the sensitive detection of single target molecules. Their hydrophilic-in-hydrophobic (HIH) nature enables the isolation and analysis of individual beads, carrying the target molecules, which can be further manipulated accurately through optical tweezer (OT) setups. However, these Teflon HIH-microwell platforms are conventionally fabricated through a complex, time-consuming and labor-intensive dry lift-off procedure which involves a series of major steps, limiting the up-scaling potential of these platforms. Alternative Teflon-based microwell fabrication methods have been extensively explored in literature but they preclude the generation of hydrophobic wells with hydrophilic bottom, thereby hampering the bioassay performance. Here, we present a new Teflon-on-glass molding method for the high throughput fabrication of hydrophilic-in-hydrophobic (HIH) microwell arrays, able to empower bead-based digital bioassays. Microwells 2.95 μm in depth and 3.86 μm in diameter were obtained to host individual beads. In these microwell arrays, sealing of reagents was demonstrated with an efficiency of 100% and seeding of superparamagnetic beads was achieved with an efficiency of 99.6%. The proposed method requires half as many steps when compared to the traditional dry lift-off process, is freely scalable and has the potential to be implemented in different bead-based bioassay applications. MDPI 2018-11-01 /pmc/articles/PMC6266592/ /pubmed/30388800 http://dx.doi.org/10.3390/ma11112154 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Tripodi, Lisa
Ven, Karen
Kil, Dries
Rutten, Iene
Puers, Robert
Lammertyn, Jeroen
Teflon-on-Glass Molding Enables High-Throughput Fabrication of Hydrophilic-in-Hydrophobic Microwells for Bead-Based Digital Bioassays
title Teflon-on-Glass Molding Enables High-Throughput Fabrication of Hydrophilic-in-Hydrophobic Microwells for Bead-Based Digital Bioassays
title_full Teflon-on-Glass Molding Enables High-Throughput Fabrication of Hydrophilic-in-Hydrophobic Microwells for Bead-Based Digital Bioassays
title_fullStr Teflon-on-Glass Molding Enables High-Throughput Fabrication of Hydrophilic-in-Hydrophobic Microwells for Bead-Based Digital Bioassays
title_full_unstemmed Teflon-on-Glass Molding Enables High-Throughput Fabrication of Hydrophilic-in-Hydrophobic Microwells for Bead-Based Digital Bioassays
title_short Teflon-on-Glass Molding Enables High-Throughput Fabrication of Hydrophilic-in-Hydrophobic Microwells for Bead-Based Digital Bioassays
title_sort teflon-on-glass molding enables high-throughput fabrication of hydrophilic-in-hydrophobic microwells for bead-based digital bioassays
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6266592/
https://www.ncbi.nlm.nih.gov/pubmed/30388800
http://dx.doi.org/10.3390/ma11112154
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