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3D printed water-soluble scaffolds for rapid production of PDMS micro-fluidic flow chambers
Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Institutionen för fysik.
Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Institutionen för fysik.ORCID-id: 0000-0001-5475-1422
Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Institutionen för fysik.
Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Institutionen för fysik.
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2018 (Engelska)Ingår i: Scientific Reports, ISSN 2045-2322, E-ISSN 2045-2322, Vol. 8, nr 1, artikel-id 3372Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

We report a novel method for fabrication of three-dimensional (3D) biocompatible micro-fluidic flow chambers in polydimethylsiloxane (PDMS) by 3D-printing water-soluble polyvinyl alcohol (PVA) filaments as master scaffolds. The scaffolds are first embedded in the PDMS and later residue-free dissolved in water leaving an inscription of the scaffolds in the hardened PDMS. We demonstrate the strength of our method using a regular, cheap 3D printer, and evaluate the inscription process and the channels micro-fluidic properties using image analysis and digital holographic microscopy. Furthermore, we provide a protocol that allows for direct printing on coverslips and we show that flow chambers with a channel cross section down to 40 x 300 μm can be realized within 60 min. These flow channels are perfectly transparent, biocompatible and can be used for microscopic applications without further treatment. Our proposed protocols facilitate an easy, fast and adaptable production of micro-fluidic channel designs that are cost-effective, do not require specialized training and can be used for a variety of cell and bacterial assays. To help readers reproduce our micro-fluidic devices, we provide: full preparation protocols, 3D-printing CAD files for channel scaffolds and our custom-made molding device, 3D printer build-plate leveling instructions, and G-code.

Ort, förlag, år, upplaga, sidor
Nature Publishing Group, 2018. Vol. 8, nr 1, artikel-id 3372
Nationell ämneskategori
Annan materialteknik Övrig annan teknik Annan fysik
Identifikatorer
URN: urn:nbn:se:umu:diva-144631DOI: 10.1038/s41598-018-21638-wISI: 000425500300044OAI: oai:DiVA.org:umu-144631DiVA, id: diva2:1181373
Forskningsfinansiär
Vetenskapsrådet, 2013-5379Kempestiftelserna, JCK-1622Tillgänglig från: 2018-02-08 Skapad: 2018-02-08 Senast uppdaterad: 2018-08-16Bibliografiskt granskad
Ingår i avhandling
1. Digital holography and image processing methods for applications in biophysics
Öppna denna publikation i ny flik eller fönster >>Digital holography and image processing methods for applications in biophysics
2018 (Engelska)Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
Abstract [en]

Understanding dynamic mechanisms, morphology and behavior of bacteria are important to develop new therapeutics to cure diseases. For example, bacterial adhesion mechanisms are prerequisites for initiation of infections and for several bacterial strains this adhesion process is mediated by adhesive surface organelles, also known as fimbriae. Escherichia coli (E. coli) is a bacterium expressing fimbriae of which pathogenic strains can cause severe diseases in fluidic environments such as the urinary tract and intestine. To better understand how E. coli cells attach and remain attached to surfaces when exposed to a fluid flow using their fimbriae, experiments using microfluidic channels are important; and to assess quantitative information of the adhesion process and cellular information of morphology, location and orientation, the imaging capability of the experimental technique is vital.

In-line digital holographic microscopy (DHM) is a powerful imaging technique that can be realized around a conventional light microscope. It is a non-invasive technique without the need of staining or sectioning of the sample to be observed in vitro. DHM provides holograms containing three-dimensional (3D) intensity and phase information of cells under study with high temporal and spatial resolution. By applying image processing algorithms to the holograms, quantitative measurements can provide information of position, shape, orientation, optical thickness of the cell, as well as dynamic cell properties such as speed, growing rate, etc.

In this thesis, we aim to improve the DHM technique and develop image processing methods to track and assess cellular properties in microfluidic channels to shed light on bacterial adhesion and cell morphology. To achieve this, we implemented a DHM technique and developed image processing algorithms to provide for a robust and quantitative analysis of holograms. We improved the cell detection accuracy and efficiency in DHM holograms by developing an algorithm for detection of cell diffraction patterns. To improve the 3D detection accuracy using in-line digital holography, we developed a novel iterative algorithm that use multiple-wavelengths. We verified our algorithms using synthetic, colloidal and cell data and applied the algorithms for detecting, tracking and analysis. We demonstrated the performance when tracking bacteria with sub-micrometer accuracy and kHz temporal resolution, as well as how DHM can be used to profile a microfluidic flow using a large number of colloidal particles. We also demonstrated how the results of cell shape analysis based on image segmentation can be used to estimate the hydrodynamic force on tethered capsule-shaped cells in micro-fluidic flows near a surface.

Ort, förlag, år, upplaga, sidor
Umeå: Umeå University, 2018. s. 59
Nyckelord
Digital holographic microscopy, image processing, image reconstruction, bacterial adhesion, cell morphology, algorithm development, software design, quantitative measurement, microfluidics, multidisciplinary research
Nationell ämneskategori
Biofysik Datorseende och robotik (autonoma system)
Forskningsämne
signalbehandling; teknisk fysik med inriktningen mikrosystemteknik
Identifikatorer
urn:nbn:se:umu:diva-150687 (URN)978-91-7601-915-3 (ISBN)
Disputation
2018-09-07, Naturvetarhuset, N430, Umeå, 13:15 (Engelska)
Opponent
Handledare
Tillgänglig från: 2018-08-17 Skapad: 2018-08-15 Senast uppdaterad: 2018-08-16Bibliografiskt granskad

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Dahlberg, TobiasStangner, TimHanqing, ZhangWiklund, KristerLundberg, PetterEdman, LudvigAndersson, Magnus

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