传感器类型
电化学生物传感器
检测对象
去甲肾上腺素(norepinephrine, NE,Krebs 溶液);神经活动/动作电位(活体豚鼠听觉皮层组织);拟检测病原体、蛋白质、DNA(未实际样品)
检测原理
硼掺杂多晶金刚石(B-doped poly-C)工作电极具有宽电位窗口和低背景电流。体外检测时,去甲肾上腺素(NE)扩散至电极表面并发生直接电化学氧化,产生法拉第电流;循环伏安法下氧化峰电流随 NE 浓度升高而增大,经 Krebs 溶液背景扣除后可在约5 nM水平区分信号。对于神经电记录,B-doped poly-C 微电极阵列置于脑组织外,记录神经元动作电位,经滤波和小波去噪提高信噪比。对于拟议病原体/蛋白/DNA 传感器,金刚石表面经 H/O/F 端基处理并接枝抗体或核酸探针,目标物结合后改变界面电化学特性,再通过三电极体系读出电流信号。该体系主要依赖直接电化学氧化与表面功能化选择性识别,未采用 HCR、RCA 或酶催化沉积等放大策略。
检测灵敏度
LOD: 500 fM(B-doped poly-C);LOD: 5 nM(NE)
效应效果
体内实验中,金刚石神经探针植入活体豚鼠听觉皮层,记录到与2 Hz听觉刺激对应的500 ms周期性峰,但信噪比低于常见硅基探针;电极阻抗约70 kΩ@1 kHz,绝缘电阻>100 kΩ,经一维小波去噪后信号改善。全金刚石SMM探针信噪比与其他金刚石电极相当但略低,主要因掺杂金刚石导电性低于金属互连。体外NE检测中,1、5、15 nM循环伏安曲线可区分,定量限约5 nM,背景电流极低。氟端基电极还原侧电位窗口更大,1 M KCl中实现4.29 V电位窗口。作者认为金刚石微加工与表面功能化可制造新一代病原体微传感器。
传感器的构成
- 基底/结构绝缘层:未掺杂多晶金刚石(undoped poly-C,10^7 Ω-cm),作为探针结构层与绝缘层。
- 换能器电极层:硼掺杂多晶金刚石(boron-doped poly-C,10^3 Ω-cm),作为工作电极、互连和电记录电极。
- 辅助电极:铂(Pt)对电极与氯化银/银(Ag/AgCl)参比电极,构成三电极电化学检测体系。
- 表面端基层:经 H2、O2、CF4 等离子处理形成氢端基、氧端基或氟端基,调节电位窗口与界面电化学性质。
- 识别功能化层:可选接枝核酸或蛋白质/抗体(nucleic acid/protein/antibody),用于选择性结合目标分子或病原体。
- 信号产生界面:硼掺杂金刚石工作电极表面直接氧化去甲肾上腺素(NE),产生与浓度相关的电化学电流。
中文摘要
金刚石因其化学惰性、生物相容性等独特性质组合而备受关注。多晶金刚石(poly-C)已被用于利用电化学方法和抗原-抗体结合检测生物分子的实验性生物传感器。硼掺杂 poly-C 电极由于具有宽电位窗口、低背景电流和噪声以及低检测限(低至500 fM),在电化学应用中表现出显著优势。poly-C 的生物相容性与钛和316不锈钢等常用植入材料相当甚至更优。我们开发了基于金刚石的神经微电极阵列(MEA),因为 poly-C 作为生物传感器材料具有吸引力。这些金刚石探针已用于体内电生理记录和体外电化学检测。poly-C 电极可用于记录神经活动。体外研究表明,金刚石探针能够在5 nM水平检测去甲肾上腺素。我们提出将金刚石微加工与表面功能化相结合,用于制造金刚石病原体微传感器。
英文摘要
Diamond is a material of interest due to its unique combination of properties, including its chemical inertness and biocompatibility. Polycrystalline diamond (poly-C) has been used in experimental biosensors that utilize electrochemical methods and antigen-antibody binding for the detection of biological molecules. Boron-doped poly-C electrodes have been found to be very advantageous for electrochemical applications due to their large potential window, low background current and noise, and low detection limits (as low as 500 fM). The biocompatibility of poly-C is found to be comparable, or superior to, other materials commonly used for implants, such as titanium and 316 stainless steel. We have developed a diamond-based, neural microelectrode-array (MEA), due to the desirability of poly-C as a biosensor. These diamond probes have been used for in vivo electrical recording and in vitro electrochemical detection. Poly-C electrodes have been used for electrical recording of neural activity. In vitro studies indicate that the diamond probe can detect norepinephrine at a 5 nM level. We propose a combination of diamond micro-machining and surface functionalization for manufacturing diamond pathogen-microsensors.