Development of a miniaturised system for the high throughput investigations of biofilm responses to antimicrobial agents.

Abstract

This thesis describes the development of a high throughput device for biofilm studies. The device generates reproducible results and can operate in several modes for different research purposes related to biofilms. The fabrication procedure is quick, easy, and cheap. The final product is robust and easy to operate. The design, production, and assembly of the device are presented, followed by physical validation of the device's performance and ability to grow E. coli biofilms. Production of a dynamic system that will aid biofilm studies with parallel readout is critical to enable the acquisition of many data points per unit of time. The performance of current systems is the main barrier to achieving high throughput studies of biofilms with reliable results. Consequently, a new device has been developed that implements 3D printing technologies. This approach has enabled the device's production with a transparent area for non-destructive biofilm observations, consisting of three parallel channels with separate growth compartments for biological repeats. The small size of the device grants extensive control over factors involved in biofilm formation and minimises the consumption of reagents. Uniformity of flow rate via separate parallel channels was achieved and provides even distribution of biofilm within the device and uniform delivery of reagents. The performance of the fabricated device was compared to static and dynamic systems widely used in biofilm research. These studies showed that the new device could be used for complex dynamic studies of biofilms and static experiments. Further validation of the fabricated device and comparing its performance against existing techniques was performed using two sterilising agents. The effect of antimicrobial properties of plant-derived compounds was studied to demonstrate the feasibility of high throughput studies using the fabricated device. The fabricated device is expected to aid biofilm studies and high throughput screening of potential antimicrobial molecules.

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Identifiers

ORCID for George Attard: orcid.org/0000-0001-8304-0742

ORCID for Jeremy Webb: orcid.org/0000-0003-2068-8589

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