Optimisation of microfluidic polymerase chain reaction devices

The invention and development of Polymerase Chain Reaction (PCR) technology have revolutionised molecular biology and molecular diagnostics. There is an urgent need to optimise the performance of these devices while reducing the total construction and operation costs. This study proposes a CFD-enabled optimisation methodology for continuous flow (CF) PCR devices with serpentine-channel structure, which enables the optimisation of DNA amplification efficiency and pressure drop to be explored while varying the width (W) and height (H) of the microfluidic (μ) channel. This is achieved by using a surrogate-enabled optimisation approach accounting for the geometrical features of a μCFPCR device by performing a series of simulations using COMSOL Multiphysics 5.4®. The values of the objectives are extracted from the CFD solutions, and the response surfaces are created using polyharmonic splines. Genetic algorithms are then used to locate the optimum design parameters. The results indicate that there is the possibility of improving the DNA concentration and the pressure drop in a PCR cycle by ~2.1 % ([W, H] = [400 μm, 50 μm]) and ~95.2 % ([W, H] = [400 μm, 80 μm]) respectively, by modifying its geometry.</jats:p

Peanut Detection Using Droplet Microfluidic Polymerase Chain Reaction Device

In this study, we integrated genetic detection for polymerase chain reaction (PCR) with microfluidics technology for the detection of peanut DNA. A cross-junction microchannel was used to induce emulsion droplets of water in oil for PCR on a chip. Compared with the single-phase flow, the emulsion droplet flow exhibited a 7.24% lower evaporation amount and prevented air bubble generation. PCR results of the droplet microfluidic PCR chip for peanut DNA fragment detection was verified by comparison with a commercial PCR thermal cycler and increased fluorescence intensity in SYBR Green reagent-based PCR. Moreover, PCR on the microfluidic PCR chip was successful for sesame, Salmonella spp., and Staphylococcus aureus. The droplet microfluidic PCR device developed in this study can be applied for peanut detection in the context of food allergy

Peanut detection using droplet microfluidic polymerase chain reaction device

[[abstract]]In this study, we integrated genetic detection for polymerase chain reaction (PCR) with microfluidics technology for the detection of peanut DNA. A cross-junction microchannel was used to induce emulsion droplets of water in oil for PCR on a chip. Compared with the single-phase flow, the emulsion droplet flow exhibited a 7.24% lower evaporation amount and prevented air bubble generation. PCR results of the droplet microfluidic PCR chip for peanut DNA fragment detection was verified by comparison with a commercial PCR thermal cycler and increased fluorescence intensity in SYBR Green reagent-based PCR. Moreover, PCR on the microfluidic PCR chip was successful for sesame, Salmonella spp., and Staphylococcus aureus. The droplet microfluidic PCR device developed in this study can be applied for peanut detection in the context of food allergy

Strategic design of flow structures for single (bio) particle analysis using droplet microfluidic platform

This thesis is designed to meet the need for knowledge of droplet-based encapsulation strategies by carrying out systematic fundamental studies based on a double-cross configuration. This configuration has been commonly used for co-encapsulation of particles or cells with multiple reagents. Particular attention is also paid to the simplicity and robustness of the channel network for real-world applications. Proposed methods of single-particle encapsulation and integration of multiple functional components are validated by the first two projects. The goal of the first project is to co-encapsulate a 1micrometer magnetic bead (MB) with multiple Quantum Dots (QDs) for further bio-decorating the QD surfaces with different molecules (i.e. single-strand DNA). The goal of the second project is to integrate into a single device a two-step reaction assay for functionalizing the surface of QDs with oligonucleotide strands while QDs are immobilized on MBs. The QD-Oligonucleotide conjugate serves as bio-sensing probes for nucleic acid detection. Additionally, to provide the fundamental knowledge for non-microfluidic-researchers to apply this approach in their single-encapsulation applications, a comprehensive experimental study is designed to span a wide range of operating parameters