Modern society, because of population ageing, suffers from a growing socio-economical impact of neurodegenerative diseases such as Alzheimer's, Parkinson's and Creutzfeldt-Jakob. Despite intense research efforts, the mechanisms responsible for the onset of these neuroinflammatory events, leading to neurodegeneration, are still unknown. Indeed, even if cytokines are well known to play a central role in the beginning of the neuroinflammatory events, their effects seem to depend on their localization and release time. Unfortunately, these local parameters, which are necessary to elucidate the neuroinflammatory mechanisms, are not accessible using traditional techniques. The aim of this project is to set-up an advanced, versatile, spatially and temporally controlled in vitro measurement tool for the electrochemical detection of signalling compounds released from a single cell. In order to reach the cellular temporal functionality and spatial dimension, we propose to use a microfluidic probe (MFP). The MFP operates by delivering a laminar stream of inducer solution through an opening (20 x 20 µm) and capturing it in a second opening, in a push-pull configuration. In the present project, microelectrodes will be added to the MFP, surrounding the apertures, in order to detect specifically the desired biomolecules in the cell micro-environment. These detections will be real-time and label-free electrochemical measurements (impedance or cyclic-voltammetry), based on the modification of the electrode surface behaviour, following biospecific interactions. The specificity of the detection will be achieved using immobilized antibodies at the electrode surface. After characterisation and improvement of the bio-sensing possibilities, the modified MFP will be used to both stimulate the cell and detect target compounds in the cell vicinity. This should allow us to characterize the intercellular signalling cascade and its kinetic properties.
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