The preparation of megaporous bodies, with potential applications in biotechnology, was attempted by following several strategies. As a first step, naive and robust scaffolds were produced by polymerization of selected monomers in the presence of a highly soluble cross-linker agent. Ion-exchange function was incorporated by particle embedding, direct chemical synthesis, or radiation-induced grafting. The total ionic capacity of such systems was 1.5 mmol H(+)/g, 1.4 mmol H(+)/g, and 17 mmol H(+)/g, respectively. These values were in agreement with the ability to bind model proteins: observed dynamic binding capacity at 50% breakthrough was ≅7.2 mg bovine serum albumin/g, ≅7.4 hen egg-white lysozyme (HEWL) mg/g, and ≅108 HEWL mg/g. In the later case, total (static) binding capacity reached 220 mg/g. It was observed that the structure and size of the megapores remained unaffected by the grafting procedure which, however, allowed for the highest protein binding capacity. Lysozyme supported on grafted body showed extensive clarification activity against a Micrococcus lysodekticus suspension in the flow-through mode, i.e., 90% destruction of suspended microbial cells was obtained with a residence time ≈ 18 min. Both protein capture and biocatalysis applications are conceivable with the 3D-megaporous materials described in this work.
Despite the widespread use of high-throughput planar patch-clamp instruments, the conventional pipette-based technique remains the method of choice for recording single-channel activity. Generally, planar platforms are not well suited for single-channel studies due to excess noise resulting from low seal resistances and the use of substrates with poor dielectric properties. Since these platforms typically use the same pore to position a cell by suction and establish a seal, biological debris from the cell suspension can contaminate the pore surface prior to seal formation, thereby reducing the seal resistance. Here, femtosecond laser ablation is used to fabricate dual-pore glass chips for use in low-noise, single-channel recordings that circumvent this problem. One pore positions a cell by suction while another nearby pore, the recording pore, avoids contamination by maintaining positive pressure until a cell is positioned and then establishes a seal. Taking advantage of the high seal resistances and low capacitive and dielectric noise realized using glass substrates, patch-clamp experiments with these dual-pore chips consistently achieved high seal resistances (>10 GU), maintained gigaseals for prolonged durations (up to 6 hrs), and enabled single-channel recordings in cell-attached mode that are comparable to those obtained by conventional patch-clamp.
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