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This paper reports on the fabrication and application of microneedles for the electroporation of adherently growing cells and intracellular recording with focus on the influence on external factors on the cell behavior. Patch-on-chip methods such as patch-clamp have been applied mostly to individual cells in suspension. However, in the human body most of cells are adherently growing cells, which motivated the development of a new chip design. The chip contains an array of 64 microneedles occupying a total area of approximately 1 mm 2. The microneedles are fabricated using dry etching of silicon, followed by an insulation, metallization and passivation. The passivation layer is opened at the tip of the needles in order to expose the metal for cell positioning via dielectrophoresis, cell electroporation, as well as intracellular recording. Various needles with diameters in the sub-micron range and heights below 10 ?m have been fabricated. Heart muscle cells, fibroblasts, and primary neuronal cells of mice were grown on these microneedle arrays. To electrically access the intracellular space, the cells were electroporated with a voltage of ±2 V. Preliminary tests show that more than 80 % of the cells could successfully be porated. ©2008 IEEE

This paper reports on the characterization of dry etching-based processes for the fabrication of silicon microneedles using a design of experiment (DoE) approach. The possibility of using such microneedles as protruding microelectrodes able to electroporate adherently growing cells and record intracellular potentials motivates the systematic analysis of the influence of etching parameters on the needle shape. Two processes are characterized: a fully isotropic etch process and a three-step etching approach. In the first case, the shape of the microneedles is defined by a single etch step. For the stepped method, the structures are realized using the following sequence: a first, isotropic step defines the tip; this is followed by anisotropic etching that increases the height of the needle; a final isotropic procedure thins the microneedle and sharpens its tip. From the various process parameters tested, it is concluded that the isotropic fabrication is influenced mostly by four process parameters, whereas six parameters dominantly govern the outcome of the stepped etching technique. The dependence of the needle shape on the etch mask diameter is also investigated. Microneedles with diameters down to the sub-micrometer range and heights below 10 μm are obtained. The experimental design is performed using the D-optimal method. The resulting geometry, i.e. heights, diameters and radii of curvature measured at different positions, is extracted from scanning electron micrographs of needle cross-sections obtained from cuts by focused ion beam. The process parameters are used as inputs and the geometry features of the microneedles as outputs for the analysis of the process.

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