Continuous and Segmented-Flow Microfluidics for Biomolecular Analysis
File(s)
chapter 3 video 3
chapter 3 video 2
Author(s)
Pereira, Fiona Marie
Type
Thesis
Abstract
Segmented flows in microfluidics have attracted much recent attention. Of particular interest is the
use of nanolitre droplets as interface tools in microfluidics. A droplet interface connects
techniques while leaving them sufficiently independent of each other; a task that difficult to
perform within a continuous flow system.
The first part of this thesis is dedicated to sample preparation and analysis in continuous flow
microfluidic systems. The polymerase chain reaction (PCR) is investigated, using electroosmotic
flow to transport the sample across various temperature zones. Additionally, a novel sieving matrix
for electrophoretic separation of dsDNA fragments and PCR amplicons on microchip and
capillary is demonstrated.
The second part of this thesis focuses on interfacing segmented and continuous flows. Two novel
interfaces are described and demonstrated. The first interface connects droplet flows and a chip-based
electrophoresis device. Using this interface, samples or reactions performed in droplets can
be directly transferred to a separation channel without suspending the separation. This allows
multiple samples to be analysed in a single separation channel, without cross contamination
between droplets. Consequently, PCR reactions and dsDNA calibration ladders can be prepared in
droplet format and analysed in high throughput. The second interface links nano-liquid
chromatography and MALDI mass spectrometry. Droplet fractionation post nano-LC separation
is used to preserve resolution between separated bands. The droplets are subsequently delivered to
a MALDI plate for mass spectrometric analysis by removing the continuous oil phase using a
hydrophobic oleophilic membrane.
The tools developed here reduce manual intervention and provide a link between multiple
analytical techniques involved in biomolecule analysis. These innovations will improve
reproducibility and reduce cross-contamination between samples.
use of nanolitre droplets as interface tools in microfluidics. A droplet interface connects
techniques while leaving them sufficiently independent of each other; a task that difficult to
perform within a continuous flow system.
The first part of this thesis is dedicated to sample preparation and analysis in continuous flow
microfluidic systems. The polymerase chain reaction (PCR) is investigated, using electroosmotic
flow to transport the sample across various temperature zones. Additionally, a novel sieving matrix
for electrophoretic separation of dsDNA fragments and PCR amplicons on microchip and
capillary is demonstrated.
The second part of this thesis focuses on interfacing segmented and continuous flows. Two novel
interfaces are described and demonstrated. The first interface connects droplet flows and a chip-based
electrophoresis device. Using this interface, samples or reactions performed in droplets can
be directly transferred to a separation channel without suspending the separation. This allows
multiple samples to be analysed in a single separation channel, without cross contamination
between droplets. Consequently, PCR reactions and dsDNA calibration ladders can be prepared in
droplet format and analysed in high throughput. The second interface links nano-liquid
chromatography and MALDI mass spectrometry. Droplet fractionation post nano-LC separation
is used to preserve resolution between separated bands. The droplets are subsequently delivered to
a MALDI plate for mass spectrometric analysis by removing the continuous oil phase using a
hydrophobic oleophilic membrane.
The tools developed here reduce manual intervention and provide a link between multiple
analytical techniques involved in biomolecule analysis. These innovations will improve
reproducibility and reduce cross-contamination between samples.
Date Issued
2011-08
Date Awarded
2012-02
Copyright Statement
Attribution NoDerivatives 4.0 International Licence (CC BY-ND)
Advisor
deMello, Andrew
Creator
Pereira, Fiona Marie
Publisher Department
Chemistry
Publisher Institution
Imperial College London
Qualification Level
Doctoral
Qualification Name
Doctor of Philosophy (PhD)