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Whether at the intramolecular or cellular scale in organisms, cell-cell adhesion adapt to external mechanical cues arising from the static environment of cells and from dynamic interactions between neighboring cells. Cell-cell adhesions need to resist detachment forces to secure the integrity and internal organization of organisms. In the past, various techniques have been developed to characterize adhesion properties of molecules and cells in vitro, and to understand how cells sense and probe their environment. Atomic force microscopy and dual-pipette aspiration, where cells are mainly present in suspension, are common methods for studying detachment forces of cell-cell adhesions. How cell-cell adhesion forces are developed for adherent and environment-adapted cells, however, is less clear. Here, we designed the Cell-Cell Separation Device (CC-SD), a microstructured substrate that measures both intercellular forces and external stresses of cells towards the matrix. The design is based on micropillar arrays originally designed for cell traction-force measurements. We designed PDMS micropillar-blocks, to which cells could adhere and be able to connect to each other across the gap. Controlled stretching of the whole substrate changed the distance between blocks and increased gap size. That allowed us to apply strains to cell-cell contacts, eventually leading to cell-cell adhesion detachment, which was measured by pillar deflections. The CC-SD provided an increase of the gap between the blocks of up to 2.4-fold, which was sufficient to separate substrate-attached cells with fully developed F-actin network. Simultaneously measured pillar deflections allowed us to address cellular response to the intercellular strain applied. The CC-SD thus opens up possibilities for the analysis of intercellular force detachments and sheds light on the robustness of cell-cell adhesions in dynamic processes in tissue development.
An electrochemical study with three redox substances on a carbon based nanogap electrode array
(2020)
Here we present the highly sensitive detection of dopamine using gold nanogap IDAs with redox-cycling amplification. Through the combination with a facile electrochemical activation and a chronoamperometric multistep protocol fouling of the gold electrode surface can be prevented and a sensitivity of 14 nA μM -1 with excellent linearity up to 10 μM is achieved. The low-cost and reproducible wafer level fabrication process of the nanogap IDAs plays a key role. Electrode and substrate materials can be nearly arbitrarily chosen. Also the gap sizes could be adjusted down to sub-100 nm dimensions with this versatile approach, allowing for very high signal amplification. In comparison to the current gold standard, fastscan cyclic voltammetry (FSCV) with carbon fiber microelectrodes (CFMEs), which suffers from high background currents, no elaborate data processing and high-end electronic equipment is needed. Employing our flexible, easy and inexpensive method, DA monitoring with a short acquisition period and a detection limit less than 200 nM is successfully demonstrated.