Aoudjane, S; (2016) Bio-analytics - from current clinical practice to all-optical microfluidic technology. Doctoral thesis , UCL (University College London).

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Abstract

There are many instances in which one would measure biological systems along a temperature gradient. This is because thermodynamics dictate the dynamics of biological processes. Here, systems are engineered where thermodynamic changes in biological systems are enacted by the use of optical heating. This optical heating allows the use of microfluidic analysis systems in making measurement of these changes. Furthermore, measurement of the temperature changes themselves aremade remotely of the sample, utilizing optical techniques. Overall these methods results in measurement of changes in biological systems, in relation to temperature change, using much lower volumes of valuable samples and reagents, than in comparable commercially available systems. Specifically the stability of a fluorescent protein was measured in a bespoke microfluidic device using all optical heating, enacted by use of a infra-red laser. By measuring the decrease in fluorescence of that protein as it became thermally denatured. This was performed using a measured volume of 15nl which is many orders of magnitude less than that used in a commercially available system. Work is ongoing, to generalize this, to nonfluorescent proteins by measuring the change in intrinsic fluorescence of proteins, as a function of temperature. A key technology in which temperature changes are enacted in a biological system is PCR, here controlled cycling of temperature is performed to induce an exponential increase in the amount of a DNA target. This process underpins sequencing of DNA as it allows enough DNA for the sequence to be detected. A microfluidic device would be constructed to perform PCR thermocycling using optical heating and thermometry. Prior to this a DNA sequencing study was conducted of the mutational patterns of HBV in a clinical scenario. This study would provide sample for, and would be used as a comparison for the construction of a microfluidic all optical qPCR device.

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