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Bacterial nano-scale cultures for rapid multiplexed antibiotic susceptibility testing
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
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2013 (English)Manuscript (preprint) (Other academic)
Place, publisher, year, edition, pages
2013.
National Category
Biomedical Laboratory Science/Technology
Identifiers
URN: urn:nbn:se:kth:diva-140111OAI: oai:DiVA.org:kth-140111DiVA: diva2:688584
Note

QS 2014

Available from: 2014-01-17 Created: 2014-01-17 Last updated: 2014-01-17Bibliographically approved
In thesis
1. Miniaturised Microwell-based Cell Assays
Open this publication in new window or tab >>Miniaturised Microwell-based Cell Assays
2014 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Cell heterogeneity in genetically identical cell populations is becoming a well-known and important phenomenon in cell biology. Current methods commonly utilise population-based analysis founded on averaged result. Hence there is a need for high-throughput cell assays on the single-cell level. By using miniaturised devices it is possible to enhance spatial and temporal control of the individual cells, increase the potential throughput and minimise the needed sample and reagent volume while enabling a wide range of biological applications.

This thesis is based on the results generated with a miniaturised microwell slide for cell assays. The microwell slide’s high-throughput compartmentalised configuration enables several hundred isolated experiments to be run simultaneously. The bottom of the wells is made out of a thin glass slide, which supports high-resolution imaging. The slide has a standardised format and its’ compatibility with conventional instruments is used extensively throughout the thesis. The presented papers all contribute to the development of the microwell slide by adding technical value or increasing the number of potential applications. For example, the slide was success-fully implemented as a chip-to-world output format for single microfluidic droplets in Paper I, by interfacing two miniaturised systems with fluorescence-activated cell sorting. In Paper II and III, microfluidic channels were integrated to increase spatial and temporal control of the added samples and reagents. In Paper II an automated stepwise concentration gradient generator was developed delivering a drug gradient to adherent mammalian cells in designated wells. In Paper III fluidic-imposed shear stress on endothelial cells was studied. In Paper IV, the slide was functionalised by coating the surfaces of the wells with several antibiotics at a defined concentration range. The coated slide was used for multiplex antibiotic susceptibility testing of bacterial pathogens, using an algorithm-based identification of the point defining lag to exponential phase transition. It successfully determined the pathogens susceptibility profile in 3-5 hours. Finally, in Paper V, a method to retrieve bacteria colonies with a desired phenotype from the wells for downstream genetic analysis was developed. In summary, the presented work has furthered the development of miniaturised high-throughput tools for various cell heterogeneity assays.

Place, publisher, year, edition, pages
Stockholm: KTH Royal Institute of Technology, 2014. iv, 70 p.
Series
TRITA-BIO-Report, ISSN 1654-2312 ; 2014:2
Keyword
microwell, miniaturisation, high-resolution imaging, high-throughput, cell culture, single-cell, clone, heterogeneity, antibiotic susceptibility testing, concentration gradient, interfacing, microchannels, cell retrieval
National Category
Biomedical Laboratory Science/Technology
Identifiers
urn:nbn:se:kth:diva-140113 (URN)978-91-7501-982-6 (ISBN)
Public defence
2014-02-07, Air & Fire - Gamma building, Tomtebodavägen 23A, Solna, 10:00 (English)
Opponent
Supervisors
Note

QC 20140117. The abstract published on January 24th, 2014.

Available from: 2014-01-17 Created: 2014-01-17 Last updated: 2014-01-24Bibliographically approved

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CiteExportLink to record
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Citation style
  • apa
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