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@PHDTHESIS{Chen:1005797,
author = {Chen, La},
title = {{D}esign, {I}mplementation and {A}pplication of {H}igh
{T}hroughput {M}agnetic {T}weezers for {C}ell {M}echanics
{S}tudies},
school = {RWTH Aachen},
type = {Dissertation},
address = {Aachen},
publisher = {RWTH Aachen},
reportid = {FZJ-2023-01642},
pages = {113 p.},
year = {2016},
note = {Dissertation, RWTH Aachen, 2016},
abstract = {Recently it has been shown that the mechanical properties
of cells play a very important role in various biological
processes. Most living cells are small, fragile and highly
heterogeneous. It is frequently observed that there is a
large inherent variation in mechanical properties from cell
to cell. Therefore, high throughput microrheology methods
are always favorable in cell mechanics studies. Furthermore,
high forces are usually needed to study cells with high
stiffness and to analyze nonlinear mechanical properties
such as stiffening or fluidization phenomena in cells.
However, most available microrheology tools are limited to
small force, poor maneuverability, and low throughput. In
this context, the current thesis presents the design and
implementation of two types of magnetic probe based
microrheometers: magnetic tweezers (MT) and magnetic
twisting cytometry (MTC), in both of which high throughput,
high force (stress), and good maneuverability were
successfully achieved at the same time. With the help of
these tools, the mechanics of rat cardiomyocytes and brain
cells were characterized. The first part of this work
focuses on the implementation of a high throughput, high
force tri-pole electromagnetic tweezers which can achieve 2D
actuation. For a given magnetic bead, the maximal force of
tri-pole magnetic tweezers depends on the size of the
workspace, the width of the magnetic tips, and on the
saturation magnetization of the tip material. In order to
conveniently calibrate and study the force behavior, an
inverse force model based on a numerical solver and active
video tracking based feedback control were implemented.
Material with high permeability was adopted as the main yoke
to reduce the coil current. The electronics and software
were custom-made to achieve high performance. For example,
with a workspace of 60×60 µm2, a force of up to 1 nN can
be applied on a 2.8 µm superparamagnetic bead in any
direction within the plane at a speed of up to 1 kHz.
However, the practically achieved saturation forces are
usually lower than predicted values, which can be ascribed
to two factors: magnetic performance deterioration near the
cutting edges of the tips and 3D geometrical effect. The
high power laser used in cutting causes segregation and
morphological roughness near the cutting edge. Moreover, the
geometry of the magnetic tips plays an important role
regarding the force behavior. In the second part, the
corrosion of the magnetic tips in several cell culture media
was characterized. Obvious accelerated corrosion was
observed in cardiomyocyte and neuronal cell media, but not
in HEK cell medium. Both the electrochemical deposition of
polypyrrole and the pyrolytical deposition of parylene-C
were examined for passivation. It was found that the quality
of polypyrrole deposition is insufficient in the area near
the edges of the tweezers tips where they had been
laser-cut. However, the parylene coating exhibits excellent
isolation properties. Both cardiomyocyte and primary
neuronal cell can be cultured on parylene-coated magnetic
parts for a long time. In addition, the coated parts can
also withstand repeated high magnetic field application. In
the third part, based on the magnetic tweezers setup, a
novel optical 2D magnetic twisting cytometry was
implemented, in which both the strength and direction of the
twisting field can be controlled. In the MTC system, both
polarization and twisting magnetic field were based on
electromagnets. A separate high field electromagnet was
utilized to magnetize the ferromagnetic particles bound on
the surface of the cells. The existing hex-pole yoke
electromagnet, but without tips, was used to apply the
twisting magnetic field. When the twisting field is less
than 100 G, good linearity and small phase error can be
achieved. Using the heterodyning technology, the measurement
frequencies were extended up to 1 kHz. In the last part of
the thesis, these developed microrheology tools were used to
study the mechanical properties of rat cardiomyocytes and
brain cells. Both the creep and the frequency response of
cardiomyocyte HL-1 cells were characterized with the
instrument operated in magnetic tweezers mode and in
magnetic twisting cytometry mode, respectively. In both
modes, the stiffness of HL-1 cells exhibits approximately
log-normal distributions. High heterogeneity of single cell
stiffness was also noticed. When HL-1 cells were cultured on
a stiff substrate, there was an obvious stiffening effect at
low frequency, which depends on the prestress generated by
myosin activity. In addition, the mechanical properties of
rat neuronal and glial cells were studied with magnetic
tweezers. It was found that with increasing maturity, the
stiffness of both neuron and glia increases. The power-law
exponent of neuronal cells decreases with increasing cell
maturity, but the one of glia cells does not change.
Especially in early stage, it was found that there is high
tension in neurites. Furthermore, the neuronal somas become
stiffer with the applied stress. Both the elastic modulus of
neurons and glia were also sensitive to the rigidity of the
substrate.},
cin = {IBI-3},
cid = {I:(DE-Juel1)IBI-3-20200312},
pnm = {5241 - Molecular Information Processing in Cellular Systems
(POF4-524)},
pid = {G:(DE-HGF)POF4-5241},
typ = {PUB:(DE-HGF)11},
url = {https://juser.fz-juelich.de/record/1005797},
}