电化学生物传感器 2009

Biotelemetric monitoring of brain neurochemistry in conscious rats using microsensors and biosensors.

Sensors (Basel, Switzerland) Calia G, Rocchitta G, Migheli R, Puggioni G, Spissu Y, Bazzu G, Mazzarello V, Lowry JP, O'Neill RD, Desole MS, Serra PA
阅读原文 PDF DOI PubMed

组成图示

Biotelemetric monitoring of brain neu... 传感器构成示意图

点击图片查看大图 · 依据论文自动绘制

传感器类型

电化学生物传感器

检测对象

抗坏血酸(ascorbic acid, AA)、氧气(oxygen, O2)、葡萄糖(glucose);样品基质为清醒大鼠纹状体细胞外液/脑组织。

检测原理

系统采用三电极安培法。AA在碳复合微电极+120 mV下被氧化为脱氢抗坏血酸,释放电子产生阳极电流;O2在碳微电极-400 mV下经两步还原生成H2O2并进一步还原为H2O,产生阴极电流。葡萄糖由Pt电极表面GOx催化氧化,生成H2O2;H2O2在+700 mV下于Pt电极氧化,产生与葡萄糖浓度成正比的阳极电流。p-OPD膜固定GOx并控制传质,提高选择性。电极电流经I/V转换、低通滤波、ADC数字化,由AM发射器无线传输至PC。信号随被测物浓度变化,AA和O2呈线性,葡萄糖在低浓度线性、高浓度呈米氏动力学。

检测灵敏度

AA: 线性范围 0–1 mM;灵敏度: 7.3 pA µM–1(植入后 2.9 pA µM–1);R^2 = 0.9959。O2: 灵敏度: 213 ± 2 pA µM–1;R^2 = 0.989;植入后灵敏度下降 18%。Glucose: 线性范围 0–2 mM;灵敏度: 15.2 ± 1.1 nA mM–1;R^2 = 0.987;Michaelis-Menten: Vmax = 89 ± 4 nA,KM = 4.8 ± 0.6 mM,R^2 = 0.989。

效应效果

遥测装置功耗约375 µW(125 µA),3 V锂纽扣电池(25 mAh)可支持1 Hz采样连续工作一周以上;接收端功耗225 mW,装置重量<10 g,约为大鼠体重3%。体外干扰测试显示,DOPAC(10 µM)、尿酸(10 µM)和多巴胺(1 µM)对AA、O2和葡萄糖传感器响应很小,AA(500 µM)对葡萄糖传感器仅产生0.73±0.2 nA响应。体内基线AA约350 µM,O2约33±14 µM,葡萄糖约492±35 µM;尾夹刺激使O2电流增加4.8 nA(约27 µM),葡萄糖先降后升。作者认为该低成本系统可稳定、可靠地用于研究药物对脑神经化学系统的影响。

传感器的构成

  • 工作电极(AA/O2):石墨/环氧树脂碳复合微电极(graphite-loaded epoxy resin, Araldite-M),直接安培氧化AA或还原O2。
  • 工作电极(葡萄糖):铂丝微电极(Pt wire, Ø125 µm),作为葡萄糖生物传感器换能器。
  • 识别/催化层:葡萄糖氧化酶(GOx)浸渍干燥于Pt表面,催化葡萄糖氧化生成H2O2。
  • 聚合物修饰层:邻苯二胺电聚合膜(p-OPD),在+700 mV电合成,固定GOx并允许H2O2氧化。
  • 选择性膜(O2):硝酸纤维素(cellulose nitrate)表面包覆,用于氧微传感器。
  • 绝缘/封装:聚四氟乙烯绝缘银丝/铂丝(Teflon-insulated Ag/Pt wire)和石英毛细管(silica capillary tube),提供机械支撑与电接触。
  • 遥测换能电子:单电源运放MCP6044、齐纳二极管ZXRE4001、微控制器PIC12F683、AM发射器RT4-433.92,将安培电流转电压并无线传输。

中文摘要

本研究报道了一种用于清醒自由大鼠脑内神经化学实时监测的植入式生物遥测系统。该系统将碳基安培微传感器和铂基葡萄糖生物传感器与微型化遥测装置连接,用于在自由运动大鼠纹状体中检测抗坏血酸(AA)、氧气(O2)和葡萄糖。微型装置包括单电源传感器驱动电路、电流-电压转换器、微控制器和微型数据发射器;氧化还原电流经PIC内置模数转换器数字化后,通过AM发射器发送至个人电脑。电子电路在体外不同条件下校准和测试,表现出高稳定性、低功耗和纳安电流范围内良好线性响应。体内结果证实了先前在有缆大鼠中记录的纹状体AA、氧气和葡萄糖动态变化。该方案基于简单且低成本组件,可作为研究不同药物对脑神经化学系统影响的快速可靠模型。

英文摘要

In this study we present the real-time monitoring of three key brain neurochemical species in conscious rats using implantable amperometric electrodes interfaced to a biotelemetric device. The new system, derived from a previous design, was coupled with carbon-based microsensors and a platinum-based biosensor for the detection of ascorbic acid (AA), O(2) and glucose in the striatum of untethered, freely-moving rats. The miniaturized device consisted of a single-supply sensor driver, a current-to-voltage converter, a microcontroller and a miniaturized data transmitter. The redox currents were digitized to digital values by means of an analog-to-digital converter integrated in a peripheral interface controller (PIC), and sent to a personal computer by means of a miniaturized AM transmitter. The electronics were calibrated and tested in vitro under different experimental conditions and exhibited high stability, low power consumption and good linear response in the nanoampere current range. The in-vivo results confirmed previously published observations on striatal AA, oxygen and glucose dynamics recorded in tethered rats. This approach, based on simple and inexpensive components, could be used as a rapid and reliable model for studying the effects of different drugs on brain neurochemical systems.

关键词

生物遥测电化学微传感器葡萄糖生物传感器纹状体抗坏血酸氧气