86 research outputs found
Sulfo-SMCC Prevents Annealing of Taxol-Stabilized Microtubules In Vitro
Microtubule structure and functions have been widely studied in vitro and in
cells. Research has shown that cysteines on tubulin play a crucial role in the
polymerization of microtubules. Here, we show that blocking sulfhydryl groups
of cysteines in taxol-stabilized polymerized microtubules with a commonly used
chemical crosslinker prevents temporal end-to-end annealing of microtubules in
vitro. This can dramatically affect the length distribution of the
microtubules. The crosslinker sulfosuccinimidyl
4-(N-maleimidomethyl)cyclohexane-1-carboxylate, sulfo-SMCC, consists of a
maleimide and an N-hydroxysuccinimide ester group to bind to sulfhydryl groups
and primary amines, respectively. Interestingly, addition of a maleimide dye
alone does not show the same interference with annealing in stabilized
microtubules. This study shows that the sulfhydryl groups of cysteines of
tubulin that are vital for the polymerization are also important for the
subsequent annealing of microtubules.Comment: 3 figure
The Filament Sensor for Near Real-Time Detection of Cytoskeletal Fiber Structures
A reliable extraction of filament data from microscopic images is of high
interest in the analysis of acto-myosin structures as early morphological
markers in mechanically guided differentiation of human mesenchymal stem cells
and the understanding of the underlying fiber arrangement processes. In this
paper, we propose the filament sensor (FS), a fast and robust processing
sequence which detects and records location, orientation, length and width for
each single filament of an image, and thus allows for the above described
analysis. The extraction of these features has previously not been possible
with existing methods. We evaluate the performance of the proposed FS in terms
of accuracy and speed in comparison to three existing methods with respect to
their limited output. Further, we provide a benchmark dataset of real cell
images along with filaments manually marked by a human expert as well as
simulated benchmark images. The FS clearly outperforms existing methods in
terms of computational runtime and filament extraction accuracy. The
implementation of the FS and the benchmark database are available as open
source.Comment: 32 pages, 21 figure
Persistence diagrams as morphological signatures of cells: A method to measure and compare cells within a population
Cell biologists study in parallel the morphology of cells with the regulation
mechanisms that modify this morphology. Such studies are complicated by the
inherent heterogeneity present in the cell population. It remains difficult to
define the morphology of a cell with parameters that can quantify this
heterogeneity, leaving the cell biologist to rely on manual inspection of cell
images. We propose an alternative to this manual inspection that is based on
topological data analysis. We characterise the shape of a cell by its contour
and nucleus. We build a filtering of the edges defining the contour using a
radial distance function initiated from the nucleus. This filtering is then
used to construct a persistence diagram that serves as a signature of the cell
shape. Two cells can then be compared by computing the Wasserstein distance
between their persistence diagrams. Given a cell population, we then compute a
distance matrix that includes all pairwise distances between its members. We
analyse this distance matrix using hierarchical clustering with different
linkage schemes and define a purity score that quantifies consistency between
those different schemes, which can then be used to assess homogeneity within
the cell population. We illustrate and validate our approach to identify
sub-populations in human mesenchymal stem cell populations.Comment: 21 pages, 7 Figure
Persistence diagrams as morphological signatures of cells:A method to measure and compare cells within a population
Cell biologists study in parallel the morphology of cells with the regulation mechanisms that modify this morphology. Such studies are complicated by the inherent heterogeneity present in the cell population. It remains difficult to define the morphology of a cell with parameters that can quantify this heterogeneity, leaving the cell biologist to rely on manual inspection of cell images. We propose an alternative to this manual inspection that is based on topological data analysis. We characterise the shape of a cell by its contour and nucleus. We build a filtering of the edges defining the contour using a radial distance function initiated from the nucleus. This filtering is then used to construct a persistence diagram that serves as a signature of the cell shape. Two cells can then be compared by computing the Wasserstein distance between their persistence diagrams. Given a cell population, we then compute a distance matrix that includes all pairwise distances between its members. We analyse this distance matrix using hierarchical clustering with different linkage schemes and define a purity score that quantifies consistency between those different schemes, which can then be used to assess homogeneity within the cell population. We illustrate and validate our approach to identify sub-populations in human mesenchymal stem cell populations
Topology counts: force distributions in circular spring networks
Filamentous polymer networks govern the mechanical properties of many
biological materials. Force distributions within these networks are typically
highly inhomogeneous and, although the importance of force distributions for
structural properties is well recognized, they are far from being understood
quantitatively. Using a combination of probabilistic and graph-theoretical
techniques we derive force distributions in a model system consisting of
ensembles of random linear spring networks on a circle. We show that
characteristic quantities, such as mean and variance of the force supported by
individual springs, can be derived explicitly in terms of only two parameters:
(i) average connectivity and (ii) number of nodes. Our analysis shows that a
classical mean-field approach fails to capture these characteristic quantities
correctly. In contrast, we demonstrate that network topology is a crucial
determinant of force distributions in an elastic spring network.Comment: 5 pages, 4 figures. Missing labels in Fig. 4 added. Reference fixe
The circular SiZer, inferred persistence of shape parameters and application to early stem cell differentiation
We generalize the SiZer of Chaudhuri and Marron (J. Amer. Statist. Assoc. 94
(1999) 807-823, Ann. Statist. 28 (2000) 408-428) for the detection of shape
parameters of densities on the real line to the case of circular data. It turns
out that only the wrapped Gaussian kernel gives a symmetric, strongly Lipschitz
semi-group satisfying "circular" causality, that is, not introducing possibly
artificial modes with increasing levels of smoothing. Some notable differences
between Euclidean and circular scale space theory are highlighted. Based on
this, we provide an asymptotic theory to make inference about the persistence
of shape features. The resulting circular mode persistence diagram is applied
to the analysis of early mechanically-induced differentiation in adult human
stem cells from their actin-myosin filament structure. As a consequence, the
circular SiZer based on the wrapped Gaussian kernel (WiZer) allows the
verification at a controlled error level of the observation reported by Zemel
et al. (Nat. Phys. 6 (2010) 468-473): Within early stem cell differentiation,
polarizations of stem cells exhibit preferred directions in three different
micro-environments.Comment: Published at http://dx.doi.org/10.3150/15-BEJ722 in the Bernoulli
(http://isi.cbs.nl/bernoulli/) by the International Statistical
Institute/Bernoulli Society (http://isi.cbs.nl/BS/bshome.htm
A Focal Adhesion Filament Cross-correlation Kit for fast, automated segmentation and correlation of focal adhesions and actin stress fibers in cells
This is the software described in our article 'A Focal Adhesion Filament Cross-correlation Kit for fast, automated segmentation and correlation of focal adhesions and actin stress fibers in cells' and the used datasets for image analysis and correlation.The zip file contains the microscopy images and segmentation and analysis.
The software itself is the executable java (.jar) file 'GUIFocalAdhesionOnly.jar
Topology determines force distributions in one-dimensional random spring networks
Networks of elastic fibers are ubiquitous in biological systems and often provide mechanical stability to cells and tissues. Fiber-reinforced materials are also common in technology. An important characteristic of such materials is their resistance to failure under load. Rupture occurs when fibers break under excessive force and when that failure propagates. Therefore, it is crucial to understand force distributions. Force distributions within such networks are typically highly inhomogeneous and are not well understood. Here we construct a simple one-dimensional model system with periodic boundary conditions by randomly placing linear springs on a circle. We consider ensembles of such networks that consist of N nodes and have an average degree of connectivity z but vary in topology. Using a graph-theoretical approach that accounts for the full topology of each network in the ensemble, we show that, surprisingly, the force distributions can be fully characterized in terms of the parameters (N,z). Despite the universal properties of such (N,z) ensembles, our analysis further reveals that a classical mean-field approach fails to capture force distributions correctly. We demonstrate that network topology is a crucial determinant of force distributions in elastic spring networks
Topology Counts: Force Distributions in Circular Spring Networks
Filamentous polymer networks govern the mechanical properties of many biological materials. Force distributions within these networks are typically highly inhomogeneous, and, although the importance of force distributions for structural properties is well recognized, they are far from being understood quantitatively. Using a combination of probabilistic and graph-theoretical techniques, we derive force distributions in a model system consisting of ensembles of random linear spring networks on a circle. We show that characteristic quantities, such as the mean and variance of the force supported by individual springs, can be derived explicitly in terms of only two parameters: (i) average connectivity and (ii) number of nodes. Our analysis shows that a classical mean-field approach fails to capture these characteristic quantities correctly. In contrast, we demonstrate that network topology is a crucial determinant of force distributions in an elastic spring network. Our results for 1D linear spring networks readily generalize to arbitrary dimensions
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