| 1. |
- Decrop, Deborah, et al.
(author)
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Single-step imprinting of femtoliter microwell arrays allows digital bioassays with attomolar limit of detection
- 2017
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In: ACS Applied Materials and Interfaces. - : American Chemical Society (ACS). - 1944-8244 .- 1944-8252. ; 9:12, s. 10418-10426
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Journal article (peer-reviewed)abstract
- Bead-based microwell array technology is growing as an ultrasensitive target detection tool. However, dissemination of the technology and its commercial use are hampered by current fabrication methods for hydrophilic-in-hydrophobic microwell arrays, which are either expensive or labour-intensive to manufacture, or which results in low bead seeding efficiencies. In this paper, we present a novel single-step manufacturing method for imprinting cheap and disposable hydrophilic-in-hydrophobic microwell arrays suitable for single-molecule detection. Single-step imprinting of hydrophilic-in-hydrophobic microwell arrays is made possible using an innovative surface energy replication approach by means of a hydrophobic thiol-ene polymer formulation. In this polymer, hydrophobic-moiety-containing monomers self-assemble against the hydrophobic surface of the imprinting stamp, which results in a hydrophobic replica surface after polymerization. After removing the stamp, hydrophilic wells are obtained with the well bottoms consisting of glass substrate. We demonstrate that the hydrophilic-in-hydrophobic imprinted microwell arrays enable successful and efficient self-assembly of individual water droplets and seeding of magnetic beads with loading efficiencies up to 96%. We also demonstrate the suitability of the microwell arrays for the isolation and detection of single-molecules achieving a limit of detection of 17.4 aM when performing a streptavidin-biotin binding assay. The ease of manufacturing demonstrated here is expected to allow translation of digital microwell array technology towards diagnostic applications.
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| 2. |
- Decrop, Deborah, et al.
(author)
-
Single-step manufacturing of femtoliter microwell arrays in a novel surface energy mimicking polymer
- 2015
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In: 18th International Conference on Solid-State Sensors, Actuators and Microsystems (IEEE TRANSDUCER 2015). - : IEEE.
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Conference paper (peer-reviewed)abstract
- We report a novel polymer material formulation and stamp-molding technique that enable rapid single-step manufacturing of hydrophilic-in-hydrophobic microwell arrays. We developed a modified thiol-ene-epoxy polymer (mOSTE+) formulation that mimics the surface energy of its mold during polymerization. The polymer inherits the surface energy from the mold through molecular self-assembly, in which functional monomers self-assemble at the interface between the liquid prepolymer and the mold surface. Combining this novel mOSTE+ material with a stamp-molding process leads to simultaneous surface energy mimicking and micro-structuring. This method was used to manufacture microwells with hydrophilic bottom and hydrophobic sidewall, depressed in a surrounding hydrophobic surface. The microwell arrays were successfully tested for the self-assembly of 62’000 femtoliter-droplets. Such femtoliter droplet arrays are useful for, e.g., digital ELISA and single cell/molecule analysis applications.
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| 3. |
- Zandi Shafagh, Reza, et al.
(author)
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Reaction Injection Molding of Hydrophilic-in-Hydrophobic Femtolitre-Well Arrays
- 2019
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In: Microsystems & Nanoengineering. - : Nature Publishing Group. - 2055-7434. ; :5
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Journal article (peer-reviewed)abstract
- Patterning of micro- and nanoscale topologies and surface properties of polymer devices is of particular importance for a broad range of life science applications, including cell-adhesion assays and highly sensitive bioassays. The manufacturing of such devices necessitates cumbersome multiple-step fabrication procedures and results in surface properties which degrade over time. This critically hinders their wide-spread dissemination. Here, we simultaneously mold and surface energy pattern microstructures in off-stoichiometric thiol-ene by area-selective monomer self-assembly in a rapid micro-reaction injection molding cycle. We replicated arrays of 1,843,650 hydrophilic-in-hydrophobic femtolitre-wells with long-term stable surface properties and magnetically trapped beads with 75% and 87.2% efficiency in single- and multiple-seeding events, respectively. These results form the basis for ultrasensitive digital biosensors, specifically, and for the fabrication of medical devices and life science research tools, generally.
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