生物电电流测量振动探头
Summary
制造，校准和使用非侵入性的振动探头来测量各种生物系统中的生物电的电流的描述。
Abstract
离子的主动运输所产生的电场，在许多生物系统中，往往成为重要的组织和器官的功能。例如，他们发挥了重要作用，直接在伤口愈合细胞迁移。在这里，我们描述了超灵敏的振动测量细胞外的电流探头的制造和使用。该探测器是一个绝缘的，削尖的一个小的铂黑尖（30-35微米），它可以检测在μA/厘米2的范围在生理盐水中的离子电流的金属丝。该探测器是由压电德尔振捣在200赫兹左右。离子电流的存在，探测器检测到其运动的极端之间的电压差。一个锁在了外来噪声放大器，过滤器，锁定探头的振动频率。数据记录到计算机中。探头校准的开始和结束在适当的盐水实验，用一腔适用于电流完全相同1.5μA/厘米2。我们描述了如何使探针，将系统设置为和校准。我们也证明了角膜测量技术，并表现出一些有代表性的成果从不同标本（角膜，皮肤，脑）。
Protocol
1。探头制造空白探头均购自世界精密仪器（elgiloy /不锈钢聚对二甲苯涂层的微电极）（见下文“特定的试剂和仪器表”）。探头切背后的提示25-30毫米，二甲苯绝缘约5毫米刮切端用手术刀（＃11刀片），以确保良好的连接。探头安装在一个黄金R30的连接器使用载银导电环氧树脂（如祭乐SL65）[*见下文注]。探头是在室温下储存在一夜之间让环氧树脂硬化。接下来，探头尖端镀有…
Discussion
我们描述了一种成本低，基本的，但高度敏感的振动探头在各种生物系统的非侵入性电流测量系统。
可能的修改
- 如果铂/铱电极（世界精密仪器;猫＃PTM23B20）是用于代替不锈钢，镀金阶段被淘汰。
应用
我们用振动探头测量电流：大鼠角膜2;大鼠晶状体3,4;小鼠皮肤5; 6 爪蟾蝌蚪;人体皮肤的7</su…
Disclosures
The authors have nothing to disclose.
Acknowledgements
我们非常感谢，组装振动探测系统的帮助下，普渡大学，瘫痪研究中心教授理查德Borgens。这项研究是由内授予美国国立卫生研究院1R01EY019101支持MZ和BR，和部分的加州再生医学RB1 – 01417研究所，美国国家科学基金会MCB – 0951199，赠款和不受限制从研究津贴，防止盲目性，加州大学戴维斯分校的眼科。
Materials
| Material Name | Type | Company | Catalogue Number | Comment |
|---|---|---|---|---|
| Eligoy-Stainless Electrode | World Precision Instruments | SSM33A70 | 76 mm, 7 MΩ, 1-2μm tip | |
| Gold R30 connector | www.vectorelect.com | R30 | Re-usable | |
| Silver-loaded epoxy | 3M | SL65 | Mix 1-part Resin with 1-part Hardener | |
| Dissecting microscope | Olympus | SZ40 | Magnification x6 to x40 | |
| Potassium dicyanoaurate (KAu(CN)2) | Sigma-Aldrich | 379867 | CAUTION: Toxic | |
| Chloroplatinic acid hydrate (H2PtCl6 x 6H2O) | Sigma-Aldrich | 520896 | CAUTION: Toxic | |
| Lead(II) acetate trihydrate (Pb(CH3CO2)2 x 3H2O) | Sigma-Aldrich | 185191 | CAUTION: Toxic | |
| Nano-Amp power source | Made in-house | – | Powered by six 1.5 V (AAA) batteries | |
| 3-dimensional micro-positioner | Line Tool Co. | Model H | ||
| Lock-in amplifier | Stanford Research Systems | SR530 | ||
| Digital I/O interface | National instruments | PCI-6220 | ||
| Shielded Connector Block with BNC connections | National instruments | BNC-2110 | ||
| Strathclyde Electrophysiology Software | University of Strathclyde Institute of Pharmacy and Biomedical Sciences, UK | WinWCP V4.1.5 | Free download from: http://spider.science.strath.ac.uk/sipbs/software_ses.htm | |
| Calibration Chamber | Made in-house | |||
| Constant Current Calibrator | Vibrating Probe Company | Powered by one 9 V (PP3) battery |
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Tags
Bioelectric CurrentVibrating ProbeActive Transport Of IonsBiological SystemsCell MigrationWound HealingUltrasensitive Vibrating ProbesExtracellular Electric CurrentsInsulated Metal WirePlatinum-black TipPhysiological SalinePiezoelectric BenderIonic CurrentsVoltage DifferenceLock-in AmplifierComputer RecordingCalibrationCornea MeasurementSkin MeasurementBrain Measurement
Cite This Article
Reid, B., Zhao, M. Measurement of Bioelectric Current with a Vibrating Probe. J. Vis. Exp. (47), e2358, doi:10.3791/2358 (2011).