Microdroplet reactors for high-throughput chemistry and biology
Author(s)
Srisa-Art, Monpichar
Type
Thesis
Abstract
Droplet-based microfluidic systems have recently been developed to overcome the problems
of slow mixing and dispersion associated with traditional microfluidic systems. By utilising
flow instabilities between two immiscible phases, droplets can be generated using normal
microfluidic formats. Further, aqueous solutions can be confined and mixed within droplets,
resulting in rapid homogenisation and no dispersion. Accordingly, droplet-based microfluidic
systems have been utilised in various applications in a high-throughput manner. However, the
techniques and methods for droplet formation, manipulation and detection have been
continuously studied and improved upon to develop, prepare, manipulate and implement
droplet systems for real-world applications. Since droplets can be controllably produced with
variable reagent compositions at high generation frequencies (1 kHz or above), on-line
detection and characterisation of every high-speed droplet is one of the most important
challenges associated with droplet analysis. The ability to extract information from each
droplet microreactor is crucial for applications in high-throughput analysis and screening. An
appropriate detection technique able to extract the vast amount of information produced in
such systems is key in unlocking the full capabilities of droplet-based.
In this work, a custom built confocal spectroscopic system was coupled with a droplet-based
microfluidic system to conduct high-sensitivity and high-throughput biological experiments.
The integration of a confocal system allows for online characterisation of individual droplets
in terms of their size, formation frequency, fluorescence intensity and population. The
combination of a droplet-based microfluidic system and the confocal detection setup has been
successfully used to demonstrate a few high-throughput chemical and biological applications.
For example, the droplet system was utilised to demonstrate high-throughput single cell
encapsulation, characterisation and quantification for the first time. In addition, highthroughput
binding assays and kinetic measurements using a well-known streptavidin-biotin
binding model and a protein-protein interaction were performed. Furthermore, a novel
approach for fluorescence lifetime imaging (FLIM) was developed and used to analyse
mixing patterns within droplets. Specifically, data from FLIM measurements were extracted
to determine spatially localised fluorescence lifetimes within droplets and thus a twodimensional
map of droplet mixing. Finally, the droplet-based microfluidic approach was
exploited to perform biological analysis at the single molecule level.
of slow mixing and dispersion associated with traditional microfluidic systems. By utilising
flow instabilities between two immiscible phases, droplets can be generated using normal
microfluidic formats. Further, aqueous solutions can be confined and mixed within droplets,
resulting in rapid homogenisation and no dispersion. Accordingly, droplet-based microfluidic
systems have been utilised in various applications in a high-throughput manner. However, the
techniques and methods for droplet formation, manipulation and detection have been
continuously studied and improved upon to develop, prepare, manipulate and implement
droplet systems for real-world applications. Since droplets can be controllably produced with
variable reagent compositions at high generation frequencies (1 kHz or above), on-line
detection and characterisation of every high-speed droplet is one of the most important
challenges associated with droplet analysis. The ability to extract information from each
droplet microreactor is crucial for applications in high-throughput analysis and screening. An
appropriate detection technique able to extract the vast amount of information produced in
such systems is key in unlocking the full capabilities of droplet-based.
In this work, a custom built confocal spectroscopic system was coupled with a droplet-based
microfluidic system to conduct high-sensitivity and high-throughput biological experiments.
The integration of a confocal system allows for online characterisation of individual droplets
in terms of their size, formation frequency, fluorescence intensity and population. The
combination of a droplet-based microfluidic system and the confocal detection setup has been
successfully used to demonstrate a few high-throughput chemical and biological applications.
For example, the droplet system was utilised to demonstrate high-throughput single cell
encapsulation, characterisation and quantification for the first time. In addition, highthroughput
binding assays and kinetic measurements using a well-known streptavidin-biotin
binding model and a protein-protein interaction were performed. Furthermore, a novel
approach for fluorescence lifetime imaging (FLIM) was developed and used to analyse
mixing patterns within droplets. Specifically, data from FLIM measurements were extracted
to determine spatially localised fluorescence lifetimes within droplets and thus a twodimensional
map of droplet mixing. Finally, the droplet-based microfluidic approach was
exploited to perform biological analysis at the single molecule level.
Date Issued
2010-03
Date Awarded
2010-04
Copyright Statement
Attribution NoDerivatives 4.0 International Licence (CC BY-ND)
Advisor
Edel, Joshua
de Mello, Andrew
Sponsor
Royal Thai Government
Creator
Srisa-Art, Monpichar
Publisher Department
Chemistry
Publisher Institution
Imperial College London
Qualification Level
Doctoral
Qualification Name
Doctor of Philosophy (PhD)