@misc{WestendorfBaeErlenkamperetal.2010, author = {Westendorf, Christian and Bae, Albert J. and Erlenkamper, Christoph and Galland, Edouard and Franck, Carl and Bodenschatz, Eberhard and Beta, Carsten}, title = {Live cell flattening}, series = {Postprints der Universit{\"a}t Potsdam : Mathematisch Naturwissenschaftliche Reihe}, journal = {Postprints der Universit{\"a}t Potsdam : Mathematisch Naturwissenschaftliche Reihe}, number = {835}, issn = {1866-8372}, doi = {10.25932/publishup-42831}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-428311}, pages = {17}, year = {2010}, abstract = {Eukaryotic cell flattening is valuable for improving microscopic observations, ranging from bright field (BF) to total internal reflection fluorescence (TIRF) microscopy. Fundamental processes, such as mitosis and in vivo actin polymerization, have been investigated using these techniques. Here, we review the well known agar overlayer protocol and the oil overlay method. In addition, we present more elaborate microfluidics-based techniques that provide us with a greater level of control. We demonstrate these techniques on the social amoebae Dictyostelium discoideum, comparing the advantages and disadvantages of each method.}, language = {en} } @article{BaeBetaBodenschatz2009, author = {Bae, Albert J. and Beta, Carsten and Bodenschatz, Eberhard}, title = {Rapid switching of chemical signals in microfluidic devices}, issn = {1473-0197}, doi = {10.1039/B905521e}, year = {2009}, abstract = {We present an analysis of concentration switching times in microfluidic devices. The limits of rapid switching are analyzed based on the theory of dispersion by Taylor and Aris and compared to both experiments and numerical simulations. We focus on switching times obtained by photo-activation of caged compounds in a micro-flow (flow photolysis). The performance of flow photolysis is compared to other switching techniques. A flow chart is provided to facilitate the application of our theoretical analysis to microfluidic switching devices.}, language = {en} } @article{WestendorfNegreteBaeetal.2013, author = {Westendorf, Christian and Negrete, Jose and Bae, Albert J. and Sandmann, Rabea and Bodenschatz, Eberhard and Beta, Carsten}, title = {Actin cytoskeleton of chemotactic amoebae operates close to the onset of oscillations}, series = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {110}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, number = {10}, publisher = {National Acad. of Sciences}, address = {Washington}, issn = {0027-8424}, doi = {10.1073/pnas.1216629110}, pages = {3853 -- 3858}, year = {2013}, abstract = {The rapid reorganization of the actin cytoskeleton in response to external stimuli is an essential property of many motile eukaryotic cells. Here, we report evidence that the actin machinery of chemotactic Dictyostelium cells operates close to an oscillatory instability. When averaging the actin response of many cells to a short pulse of the chemoattractant cAMP, we observed a transient accumulation of cortical actin reminiscent of a damped oscillation. At the single-cell level, however, the response dynamics ranged from short, strongly damped responses to slowly decaying, weakly damped oscillations. Furthermore, in a small subpopulation, we observed self-sustained oscillations in the cortical F-actin concentration. To substantiate that an oscillatory mechanism governs the actin dynamics in these cells, we systematically exposed a large number of cells to periodic pulse trains of different frequencies. Our results indicate a resonance peak at a stimulation period of around 20 s. We propose a delayed feedback model that explains our experimental findings based on a time-delay in the regulatory network of the actin system. To test the model, we performed stimulation experiments with cells that express GFP-tagged fusion proteins of Coronin and actin-interacting protein 1, as well as knockout mutants that lack Coronin and actin-interacting protein 1. These actin-binding proteins enhance the disassembly of actin filaments and thus allow us to estimate the delay time in the regulatory feedback loop. Based on this independent estimate, our model predicts an intrinsic period of 20 s, which agrees with the resonance observed in our periodic stimulation experiments.}, language = {en} } @article{BoedekerBetaFranketal.2010, author = {Boedeker, Hendrik Ulrich and Beta, Carsten and Frank, Till D. and Bodenschatz, Eberhard}, title = {Quantitative analysis of random ameboid motion}, issn = {0295-5075}, doi = {10.1209/0295-5075/90/28005}, year = {2010}, abstract = {We quantify random migration of the social ameba Dictyostelium discoideum. We demonstrate that the statistics of cell motion can be described by an underlying Langevin-type stochastic differential equation. An analytic expression for the velocity distribution function is derived. The separation into deterministic and stochastic parts of the movement shows that the cells undergo a damped motion with multiplicative noise. Both contributions to the dynamics display a distinct response to external physiological stimuli. The deterministic component depends on the developmental state and ambient levels of signaling substances, while the stochastic part does not.}, language = {en} } @article{AmselemThevesBaeetal.2012, author = {Amselem, Gabriel and Theves, Matthias and Bae, Albert J. and Bodenschatz, Eberhard and Beta, Carsten}, title = {A stochastic description of dictyostelium chemotaxis}, series = {PLoS one}, volume = {7}, journal = {PLoS one}, number = {5}, publisher = {PLoS}, address = {San Fransisco}, issn = {1932-6203}, doi = {10.1371/journal.pone.0037213}, pages = {11}, year = {2012}, abstract = {Chemotaxis, the directed motion of a cell toward a chemical source, plays a key role in many essential biological processes. Here, we derive a statistical model that quantitatively describes the chemotactic motion of eukaryotic cells in a chemical gradient. Our model is based on observations of the chemotactic motion of the social ameba Dictyostelium discoideum, a model organism for eukaryotic chemotaxis. A large number of cell trajectories in stationary, linear chemoattractant gradients is measured, using microfluidic tools in combination with automated cell tracking. We describe the directional motion as the interplay between deterministic and stochastic contributions based on a Langevin equation. The functional form of this equation is directly extracted from experimental data by angle-resolved conditional averages. It contains quadratic deterministic damping and multiplicative noise. In the presence of an external gradient, the deterministic part shows a clear angular dependence that takes the form of a force pointing in gradient direction. With increasing gradient steepness, this force passes through a maximum that coincides with maxima in both speed and directionality of the cells. The stochastic part, on the other hand, does not depend on the orientation of the directional cue and remains independent of the gradient magnitude. Numerical simulations of our probabilistic model yield quantitative agreement with the experimental distribution functions. Thus our model captures well the dynamics of chemotactic cells and can serve to quantify differences and similarities of different chemotactic eukaryotes. Finally, on the basis of our model, we can characterize the heterogeneity within a population of chemotactic cells.}, language = {en} } @article{SchaeferWestendorfBodenschatzetal.2011, author = {Schaefer, Edith and Westendorf, Christian and Bodenschatz, Eberhard and Beta, Carsten and Geil, Burkhard and Janshoff, Andreas}, title = {Shape oscillations of dictyostelium discoideum cells on ultramicroelectrodes monitored by impedance analysis}, series = {Small}, volume = {7}, journal = {Small}, number = {6}, publisher = {Wiley-Blackwell}, address = {Malden}, issn = {1613-6810}, doi = {10.1002/smll.201001955}, pages = {723 -- 726}, year = {2011}, language = {en} } @article{AmselemThevesBaeetal.2012, author = {Amselem, Gabriel and Theves, Matthias and Bae, Albert J. and Beta, Carsten and Bodenschatz, Eberhard}, title = {Control parameter description of eukaryotic chemotaxis}, series = {Physical review letters}, volume = {109}, journal = {Physical review letters}, number = {10}, publisher = {American Physical Society}, address = {College Park}, issn = {0031-9007}, doi = {10.1103/PhysRevLett.109.108103}, pages = {5}, year = {2012}, abstract = {The chemotaxis of eukaryotic cells depends both on the average concentration of the chemoattractant and on the steepness of its gradient. For the social amoeba Dictyostelium discoideum, we test quantitatively the prediction by Ueda and Shibata [Biophys. J. 93, 11 (2007)] that the efficacy of chemotaxis depends on a single control parameter only, namely, the signal-to-noise ratio (SNR), determined by the stochastic fluctuations of (i) the binding of the chemoattractant molecule to the transmembrane receptor and (ii) the intracellular activation of the effector of the signaling cascade. For SNR less than or similar to 1, the theory captures the experimental findings well, while for larger SNR noise sources further downstream in the signaling pathway need to be taken into account.}, language = {en} } @misc{BetaBodenschatz2011, author = {Beta, Carsten and Bodenschatz, Eberhard}, title = {Microfluidic tools for quantitative studies of eukaryotic chemotaxis}, series = {European journal of cell biology}, volume = {90}, journal = {European journal of cell biology}, number = {10}, publisher = {Elsevier}, address = {Jena}, issn = {0171-9335}, doi = {10.1016/j.ejcb.2011.05.006}, pages = {811 -- 816}, year = {2011}, abstract = {Over the past decade, microfluidic techniques have been established as a versatile platform to perform live cell experiments under well-controlled conditions. To investigate the directional responses of cells, stable concentration profiles of chemotactic factors can be generated in microfluidic gradient mixers that provide a high degree of spatial control. However, the times for built-up and switching of gradient profiles are in general too slow to resolve the intracellular protein translocation events of directional sensing of eukaryotes. Here, we review an example of a conventional microfluidic gradient mixer as well as the novel flow photolysis technique that achieves an increased temporal resolution by combining the photo-activation of caged compounds with the advantages of microfluidic chambers.}, language = {en} } @article{NegretePumirHsuetal.2016, author = {Negrete, Jose and Pumir, Alain and Hsu, Hsin-Fang and Westendorf, Christian and Tarantola, Marco and Beta, Carsten and Bodenschatz, Eberhard}, title = {Noisy Oscillations in the Actin Cytoskeleton of Chemotactic Amoeba}, series = {Physical review letters}, volume = {117}, journal = {Physical review letters}, publisher = {American Physical Society}, address = {College Park}, issn = {0031-9007}, doi = {10.1103/PhysRevLett.117.148102}, pages = {5}, year = {2016}, abstract = {Biological systems with their complex biochemical networks are known to be intrinsically noisy. Here we investigate the dynamics of actin polymerization of amoeboid cells, which are close to the onset of oscillations. We show that the large phenotypic variability in the polymerization dynamics can be accurately captured by a generic nonlinear oscillator model in the presence of noise. We determine the relative role of the noise with a single dimensionless, experimentally accessible parameter, thus providing a quantitative description of the variability in a population of cells. Our approach, which rests on a generic description of a system close to a Hopf bifurcation and includes the effect of noise, can characterize the dynamics of a large class of noisy systems close to an oscillatory instability.}, language = {en} }