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Title: Design, test and biological validation of microfluidic systems for blood plasma separation
Author: Kersaudy-Kerhoas, Maiwenn
ISNI:       0000 0004 2711 7479
Awarding Body: Heriot-Watt University
Current Institution: Heriot-Watt University
Date of Award: 2010
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Sample preparation has been described as the weak link in microfluidics. In particular, plasma has to be extracted from whole blood for many analysis including protein analysis, cell-free DNA detection for prenatal diagnosis and transplant monitoring. The lack of suitable devices to perform the separation at the microscale means that Lab On Chip (LOC) modules cannot be fully operated without sample preparation in a full-scale laboratory. In order to address this issue, blood flow in microchannels has been studied, and red blood cells behaviours in different geometrical environments have been classified. Several designs have been subsequently proposed to exploit some natural properties of blood flow and extract pure plasma without disturbing the cells. Furthermore, a high-level modelling method was developed to predict the behaviour of passive microfluidic networks. Additionally, the technique proposed provides useful guidance over the use of systems in more complex external environments. Experimental results have shown that plasma could be separated from undiluted whole blood in 10μm width microchannels at a flow rate of 2mL/hr. Using slightly larger structures (20μm) suitable for mass-manufacturing, diluted blood can be separated with 100% purity efficiency at high flow rate. An extensive biological validation of the extracted plasma was carried out to demonstrate its quality. To this effect Polymerase Chain Reaction (PCR) was performed to amplify targeted human genomic sequence from cell-free DNA present in the plasma. Furthermore, the influence of the sample dilution and separation efficiency on the amplification was characterised. It was shown that the sample dilution does have an influence on the amplification of house-keeping gene, but that amplification can be achieved even on high diluted samples. Additionally amplification can also be obtained on plasma samples with a range of separation efficiencies from 100% to 84%. In particular, two main points have been demonstrated (i) the extraction of plasma using combination of constrictions and bifurcations, (ii) the biological validation of the extracted plasma.
Supervisor: Dhariwal, Resh ; Desmulliez, Marc Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available