David Selmeczi

Phase transition in the collective migration of tissue cells: Experiment and model

We have recorded the swarming-like collective migration of a large number of keratocytes (tissue cells obtained from the scales of goldfish) using long-term videomicroscopy. By increasing the overall density of the migrating cells, we have been able to demonstrate experimentally a kinetic phase transition from a disordered into an ordered state. Near the critical density a complex picture emerges with interacting clusters of cells moving in groups. Motivated by these experiments we have constructed a flocking model that exhibits a continuous transition to the ordered phase, while assuming only short-range interactions and no explicit information about the knowledge of the directions of motion of neighbors. Placing cells in microfabricated arenas we found spectacular whirling behavior which we could also reproduce in simulations.

Monitoring of living cell attachment and spreading using reverse symmetry waveguide sensing

The effect of the attachment and spreading of living cells on the modes of a grating coupled reverse symmetry waveguide sensor is investigated in real time. The reverse symmetry design has an increased probing depth into the sample making it well suited for the monitoring of cell morphology. As a result, significant changes in the incoupling peak height and peak shape were observed during cell attachment and spreading. It is suggested that the area under the incoupling peaks reflects the initial cell attachment process, while the mean peak position is mostly governed by the spreading of the cells.

Injection molded chips with integrated conducting polymer electrodes for electroporation of cells

We present the design-concept for an all polymer injection molded single use microfluidic device. The fabricated devices comprise integrated conducting polymer electrodes and Luer fitting ports to allow for liquid and electrical access. A case study of low voltage electroporation of biological cells in suspension is presented. The working principle of the electroporation device is based on a focusing of the electric field by means of a constriction in the flow channel for the cells. We demonstrate the use of AC voltage for electroporation by applying a 1 kHz, ±50 V square pulse train to the electrodes and show delivery of polynucleotide fluorescent dye in 46% of human acute monocytic leukemia cells passing the constriction.

Atomic force microscopy of height fluctuations of fibroblast cells

We investigated the nanometer scale height fluctuations of 3T3 fibroblast cells with the atomic force microscope under physiological conditions. A correlation between these fluctuations and lateral cellular motility can be observed. Fluctuations measured on leading edges appear to be predominantly related to actin polymerization-depolymerization processes. We found fast (5 Hz) pulsatory behavior with 1-2 nm amplitude on a cell with low motility showing emphasized structure of stress fibers. Myosin driven contractions of stress fibers are thought to induce this pulsation.

Fast prototyping of injection molded polymer microfluidic chips

We present fast prototyping of injection molding tools by the definition of microfluidic structures in a light-curable epoxy (SU-8) directly on planar nickel mold inserts. Optimized prototype mold structures could withstand injection molding of more than 300 replicas in cyclic olefin copolymer (COC) without any signs of failure or release. The key parameters to avoid mold failure are maximum adhesion strength of the epoxy to the nickel insert and minimum interfacial energy of the epoxy pattern to the molded polymer. Optimal molding of microstructures with vertical sidewalls was found for nickel inserts pre-coated by silicon oxide before applying the structured epoxy, followed by coating of the epoxy by a fluorocarbon layer prior to injection molding. Further improvements in the mold stability were observed after homogeneous coating of the patterned epoxy by a second reflowed layer of epoxy, likely due to the resulting reduction in sidewall steepness. We employed the latter method for injection molding bondable polymer microfluidic chips with integrated conducting polymer electrode arrays that permitted the culture and on-chip analysis of cell spreading by impedance spectroscopy.

Efficient large volume electroporation of dendritic cells through micrometer scale manipulation of flow in a disposable polymer chip

We present a hybrid chip of polymer and stainless steel designed for high-throughput continuous electroporation of cells in suspension. The chip is constructed with two parallel stainless steel mesh electrodes oriented perpendicular to the liquid flow. The relatively high hydrodynamic resistance of the micrometer sized holes in the meshes compared to the main channel enforces an almost homogeneous flow velocity between the meshes. Thereby, very uniform electroporation of the cells can be accomplished. Successful electroporation of 20 million human dendritic cells with mRNA is demonstrated. The performance of the chip is similar to that of the traditional electroporation cuvette, but without an upper limit on the number of cells to be electroporated. The device is constructed with two female Luer parts and can easily be integrated with other microfluidic components. Furthermore it is fabricated from injection molded polymer parts and commercially available stainless steel mesh, making it suitable for inexpensive mass production.

Morphological changes in living cell cultures following α-particle irradiation studied by optical and atomic force microscopy

The shape of the cells and the released material (traces) from migrating adherent cells depends on the cell type, the surface coating of the culture plate, the extracellular matrix, and some other parameters. In our studies we investigated the effect of α-exposure (7-28 particle/cell/h) on cell cultures. C6 glioma cells and 3T3 cell-lines were grown on a 20 μm thick, chemically modified polypropylene foil and exposed to α-particles of an 241 Am (37 kBq) source through the foil. The average energy of the a particles was calculated by Monte-Carlo simulation of the whole collision cascade through the given pathway in air and in the foil. The α-dose was measured by CR-39 track detectors. Morphological changes of the cells were investigated by phase-contrast optical microscopy and by atomic force microscopy in low force contact mode. The static traces give information about the dynamics of cell migration and about the quality of the extracellular matrix.

Brownian Motion after Einstein: Some New Applications and New Experiments

The first half of this chapter describes the development in mathematical models of Brownian motion after Einsteins seminal papers and current applications to optical tweezers. This instrument of choice among single-molecule biophysicists is also an instrument of precision that requires an understanding of Brownian motion beyond Einsteins. This is illustrated with some applications, current and potential, and it is shown how addition of a controlled forced motion on the nano-scale of the tweezed objects thermal motion can improve the calibration of the instrument in general, and make it possible also in complex surroundings. The second half of the present chapter, starting with Sect. 9, describes the co-evolution of biological motility models with models of Brownian motion, including very recent results for how to derive cell-type-specific motility models from experimental cell trajectories.