eprintid: 19990
rev_number: 14
eprint_status: archive
userid: 2257
dir: disk0/00/01/99/90
datestamp: 2015-12-18 11:33:10
lastmod: 2016-03-07 12:31:57
status_changed: 2015-12-18 11:33:10
type: doctoralThesis
metadata_visibility: show
creators_name: Janiesch, Jan-Willi
title: Development of Droplet-Based Microfluidics for Synthetic Biology
Applications
subjects: 540
subjects: 570
subjects: 600
divisions: 120300
adv_faculty: af-12
abstract: Microfluidics combines principles of science and technology, and enables the user to handle,
process and manipulate fluids of very small volumes. This technology permits the
integration of multiple laboratory applications into one single microfabricated chip, requires
minimal manual user intervention and sample consumption, and allows enhanced
data analysis speed and precision. Due to these numerous advantages, the potential for
this technology to be applied in fundamental biophysical and biomedical research is vast.
The major aim of this thesis was to explore the capacities of microfluidics, particularly
droplet-based microfluidic technology in the following topics: 1) Mimicry of the immune
system cellular environment, with the ultimate goal of programing T cells for adoptive
T cell therapy; 2) Bottom-up assembly of minimal synthetic cells. Towards this end, a
novel approach to form gold-nanostructured and specifically biofunctionalized water-in-oil
droplets was developed. This thesis highlights the advanced properties of nanostructured
droplets to serve as 3D antigen presenting cell (APC) surrogates for T-cell stimulation.
The combination of flexible biofunctionalization and pliable physical droplet properties
work in tandem, providing a flexible and modular system that closely models in situ
APC-T cell interactions. The research within this thesis focused also on the dissection of
complex cellular sensory machinery implementing an automated droplet-based microfluidic
approach. Towards this goal, nanostructured droplets as cell-sized compartments and
droplet-based pico-injection technology were used to achieve the bottom-up assembly of
the minimal number of proteins required for a “simple synthetic cell.” While the applied
methodology has a potential for assembly of a wide range of subcellular functional units,
the focus in this thesis was on the reconstitution of the actomyosin cortex. Successful
optimization of the biochemical and biophysical conditions within the droplets allowed to
achieve precise control over the actin polymerization and actomyosin network organization
by their linkage to the droplets periphery. These experimental steps were also necessary
to generate signaling events including myosin-driven droplet migration and self-propulsion
with reduced molecular complexity compared to living cells.
date: 2015
id_scheme: DOI
id_number: 10.11588/heidok.00019990
ppn_swb: 1655516752
own_urn: urn:nbn:de:bsz:16-heidok-199904
date_accepted: 2015-12-14
advisor: HASH(0x556120af1e30)
language: eng
bibsort: JANIESCHJADEVELOPMEN2015
full_text_status: public
place_of_pub: Heidelberg
citation:   Janiesch, Jan-Willi  (2015) Development of Droplet-Based Microfluidics for Synthetic Biology Applications.  [Dissertation]     
document_url: https://archiv.ub.uni-heidelberg.de/volltextserver/19990/1/Janieschthesis.pdf