Lyon Institute of Nanotechnologies, CNRS UMR 5270, INSA de Lyon, 7, avenue Jean Capelle, 69621 Villeurbanne Cedex, France; INSERM U1028, CNRS UMR 5292, Lyon Neuroscience Research Center, Team WAKING, 8 avenue Rockefeller, University Claude Bernard Lyon 1, 69373 Lyon Cedex 08, France.
INSERM U1028, CNRS UMR 5292, Lyon Neuroscience Research Center, Team WAKING, 8 avenue Rockefeller, University Claude Bernard Lyon 1, 69373 Lyon Cedex 08, France.
Biosens Bioelectron. 2015 Oct 15;72:148-55. doi: 10.1016/j.bios.2015.05.004. Epub 2015 May 6.
Simultaneous monitoring of glucose and lactate is an important challenge for understanding brain energetics in physiological or pathological states. We demonstrate here a versatile method based on a minimally invasive single implantation in the rat brain. A silicon/SU8-polymer multi-sensing needle-shaped biosensor, was fabricated and tested. The multi-electrode array design comprises three platinum planar microelectrodes with a surface area of 40 × 200 µm(2) and a spacing of 200 µm, which were micromachined on a single 3mm long micro-needle having a 100 × 50 µm(2) cross-section for reduced tissue damage during implantation. Platinum micro-electrodes were aligned at the bottom of micro-wells obtained by photolithography on a SU8 photoresist layer. After clean room processing, each micro-electrode was functionalized inside the micro-wells by means of a micro-dispensing device, either with glucose oxidase or with lactate oxidase, which were cross-linked on the platinum electrodes. The third electrode covered with Bovine Serum Albumin (BSA) was used for the control of non-specific currents. The thick SU8 photoresist layer has revealed excellent electrical insulation of the micro-electrodes and between interconnection lines, and ensured a precise localization and packaging of the sensing enzymes on platinum micro-electrodes. During in vitro calibration with concentrations of analytes in the mM range, the micro-wells patterned in the SU8 photoresist proved to be highly effective in eliminating cross-talk signals, caused by H2O2 diffusion from closely spaced micro-electrodes. Moreover, our biosensor was successfully assayed in the rat cortex for simultaneous monitoring of both glucose and lactate during insulin and glucose administration.
同时监测葡萄糖和乳酸对于理解生理或病理状态下的大脑能量代谢是一个重要的挑战。我们在这里展示了一种基于大鼠脑内微创单一植入的多功能方法。我们制作并测试了一种硅/SU8 聚合物多传感针状生物传感器。多电极阵列设计包括三个铂平面微电极,其表面积为 40×200 µm²,间距为 200 µm,这些微电极被微加工在一个长 3mm、截面为 100×50 µm²的微针上,以减少植入过程中的组织损伤。铂微电极在 SU8 光刻胶层上的光刻微井底部对齐。经过洁净室处理后,每个微电极都通过微分配装置在微井内进行功能化,要么是葡萄糖氧化酶,要么是乳酸氧化酶,它们在铂电极上交联。覆盖牛血清白蛋白 (BSA) 的第三电极用于控制非特异性电流。厚的 SU8 光刻胶层具有优异的微电极间和互连线间的电绝缘性能,并确保了传感酶在铂微电极上的精确定位和封装。在体外使用 mM 浓度的分析物进行校准期间,SU8 光刻胶中图案化的微井被证明在消除由紧密间隔的微电极扩散引起的串扰信号方面非常有效。此外,我们的生物传感器还成功地在大鼠皮层中进行了测试,用于在胰岛素和葡萄糖给药期间同时监测葡萄糖和乳酸。