Articles by Jan Steinkühler in JoVE
Phase Behavior of Charged Vesicles Under Symmetric and Asymmetric Solution Conditions Monitored with Fluorescence Microscopy Bastian Kubsch1, Tom Robinson1, Jan Steinkühler1, Rumiana Dimova1 1Max Planck Institute of Colloids and Interfaces Experiments on phase separated giant unilamellar vesicles (GUVs) frequently neglect physiological solution conditions. This work presents approaches to study the effect of high-salinity buffer on liquid-liquid phase separation in charged multicomponent GUVs as a function of trans-membrane solution asymmetry and temperature.
Other articles by Jan Steinkühler on PubMed
From Beetles in Nature to the Laboratory: Actuating Underwater Locomotion on Hydrophobic Surfaces Langmuir : the ACS Journal of Surfaces and Colloids. Dec, 2015 | Pubmed ID: 26633751 The controlled wetting and dewetting of surfaces is a primary mechanism used by beetles in nature, such as the ladybird and the leaf beetle for underwater locomotion.1 Their adhesion to surfaces underwater is enabled through the attachment of bubbles trapped in their setae-covered legs. Locomotion, however, is performed by applying mechanical forces in order to move, attach, and detach the bubbles in a controlled manner. Under synthetic conditions, however, when a bubble is bound to a surface, it is nearly impossible to maneuver without the use of external stimuli. Thus, actuated wetting and dewetting of surfaces remain challenges. Here, electrowetting-on-dielectric (EWOD) is used for the manipulation of bubble-particle complexes on unpatterned surfaces. Bubbles nucleate on catalytic Janus disks adjacent to a hydrophobic surface. By changing the wettability of the surface through electrowetting, the bubbles show a variety of reactions, depending on the shape and periodicity of the electrical signal. Time-resolved (μs) imaging of bubble radial oscillations reveals possible mechanisms for the lateral mobility of bubbles on a surface under electrowetting: bubble instability is induced when electric pulses are carefully adjusted. This instability is used to control the surface-bound bubble locomotion and is described in terms of the change in surface energy. It is shown that a deterministic force applied normal can lead to a random walk of micrometer-sized bubbles by exploiting the phenomenon of contact angle hysteresis. Finally, bubble use in nature for underwater locomotion and the actuated bubble locomotion presented in this study are compared.
Posing for a Picture: Vesicle Immobilization in Agarose Gel Scientific Reports. 05, 2016 | Pubmed ID: 27140695 Taking a photo typically requires the object of interest to stand still. In science, imaging is potentiated by optical and electron microscopy. However, living and soft matter are not still. Thus, biological preparations for microscopy usually include a fixation step. Similarly, immobilization strategies are required for or substantially facilitate imaging of cells or lipid vesicles, and even more so for acquiring high-quality data via fluorescence-based techniques. Here, we describe a simple yet efficient method to immobilize objects such as lipid vesicles with sizes between 0.1 and 100 μm using agarose gel. We show that while large and giant unilamellar vesicles (LUVs and GUVs) can be caged in the pockets of the gel meshwork, small molecules, proteins and micelles remain free to diffuse through the gel and interact with membranes as in agarose-free solutions, and complex biochemical reactions involving several proteins can proceed in the gel. At the same time, immobilization in agarose has no adverse effect on the GUV size and stability. By applying techniques such as FRAP and FCS, we show that the lateral diffusion of lipids is not affected by the gel. Finally, our immobilization strategy allows capturing high-resolution 3D images of GUVs.
Modulating Vesicle Adhesion by Electric Fields Biophysical Journal. Oct, 2016 | Pubmed ID: 27705768 We introduce an experimental setup for modulating adhesion of giant unilamellar vesicles to a planar substrate. Adhesion is induced by the application of an external potential to a transparent indium tin oxide-coated electrode (the substrate), which enables single-vesicle studies. We demonstrate tunable and reversible adhesion of negatively charged vesicles. The adhesion energy at different potentials is calculated from the vesicle shape assessed with confocal microscopy. Two approaches for these estimates are employed: one based on the whole contour of the vesicle and a second based on the contact curvature of the membrane in the vicinity of the substrate. Both approaches agree well with each other and show that the adhering vesicles are in the weak adhesion regime for the range of explored external potentials. Using fluorescence quenching assays, we detect that, in the adhering membrane segment, only the outer bilayer leaflet of the vesicle is depleted of negatively charged fluorescent lipids, while the inner leaflet remains unaffected. We show that depletion of negatively charged lipids is consistent Poisson-Boltzmann theory, taking into account charge regulation from lipid mobility. Finally, we also show that lipid diffusion is not significantly affected in the adhering membrane segment. We believe that the approaches introduced here for modulating and assessing vesicle adhesion have many potential applications in the field of single-vesicle studies and research on membrane adhesion.