University of Twente Student Theses


Oxygen sensing in a 3D cell culture using an oxygen-sensitive dye based on a Ru(II) complex

Pouw, Fienie (2022) Oxygen sensing in a 3D cell culture using an oxygen-sensitive dye based on a Ru(II) complex.

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Abstract:While oxygen plays a major role in cell viability and function it is often neglected and/or underestimated in cell or tissue based studies. 3D oxygen mapping makes it possible to map the oxygen concentration and measure metabolic cell activity more accurately. In this study, the applicability of a Ru(II) complex based dye, [Ru(BPhen)2]2 (BPhen=batophenanthroline) with a terminated carboxylic acid group (Ru-complex), for oxygen mapping in a 3D cell/cluster-laden collagen hydrogel was validated based on optical properties, binding efficiency and toxicity. The Ru(II) based dye quenches linearly by increasing oxygen concentration (when dissolved in Milli-Q with a small percentage of DMSO). The carboxylic acid group of the Ru-complex can be covalently attached to collagen type I through 1-Ethyl-3- [3-dimethylaminopropyl]carbodiimide hydrochloride (EDC) chemistry. Optimalisation of the coupling protocol can increase binding efficiency and make it more time efficient. Unattached dye is toxic to MCF-7 cells (common breast cancer lineage) when used in a cell- and dye- laden collagen matrix, which was more significant upon excitation of the dye, which makes it unsuitable for cell applications. Additionally, the dye was insoluble in aqueous media and precipitation was observed when phosphate buffered saline (PBS) buffer or N Hydroxysulfosuccinimide (Sulfo-NHS) was used in combination with the dye. In future studies, a dye with for example a negative charge could be considered to decrease toxicity and increase solubility
Item Type:Essay (Bachelor)
Faculty:TNW: Science and Technology
Subject:35 chemistry
Programme:Biomedical Technology BSc (56226)
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