双电极电压钳（TEVC）离子通道功能检测服务Two-Electrode Voltage Clamp (TEVC) ion channel assay

以爪蟾卵母细胞为异源表达系统，通过 TEVC 技术精准测定离子通道、受体与转运蛋白的电流特性、电压依赖性和药理学响应，是国际公认的离子通道研究金标准方法。Using the Xenopus oocyte heterologous expression system, TEVC precisely measures currents, voltage dependence and pharmacological responses of ion channels, receptors and transporters — the internationally recognised gold-standard for ion channel research.

TEVC 技术原理TEVC principle

双电极电压钳（Two-Electrode Voltage Clamp，TEVC）通过两个独立的玻璃微电极同步工作：电压电极实时监测膜电位，电流电极向细胞注入补偿电流使膜电位钳制在目标值。在精准的电压控制下，跨膜离子电流得以精确测量，信噪比远优于哺乳动物细胞系统。TEVC uses two independent glass microelectrodes: a voltage electrode monitors membrane potential in real time, while a current electrode injects compensatory current to clamp the membrane at the target voltage. Under precise voltage control, transmembrane ionic currents are measured with signal-to-noise ratios far superior to mammalian cell systems.

爪蟾卵母细胞（直径 1–1.3 mm）体积大、内源通道背景低，是 TEVC 研究的理想宿主细胞，已被用于表达来自植物、动物、微生物的数千种离子通道与转运蛋白。Xenopus oocytes (diameter 1–1.3 mm) are large, with low endogenous channel background — the ideal TEVC host cell, used to express thousands of channels and transporters from plants, animals and microorganisms.

核心检测维度Core analysis dimensions

三大分析方向，覆盖通道全特征Three analysis directions for complete channel characterization

电流 – 电压关系（I-V）Current – voltage relationship (I-V)

阶梯电压方案（voltage-step protocol）测定稳态 I-V 曲线Step voltage protocol for steady-state I-V curve
揭示通道电压依赖性与门控特性Reveals voltage dependence and gating properties
通过零电流电位确定离子选择性（reversal potential）Ion selectivity via reversal potential determination
计算电导（G-V）曲线与电导密度G-V conductance curve and conductance density

动力学参数分析Kinetics analysis

激活时间常数（τ_act）：通道从关闭到开放的速率Activation time constant (τ_act): rate of channel opening
失活时间常数（τ_inact）：持续刺激下通道的关闭速率Inactivation time constant (τ_inact): channel closure under sustained stimulation
失活后恢复时间（τ_rec）Recovery from inactivation (τ_rec)
多指数拟合（单 / 双 / 三指数模型）Multi-exponential curve fitting (mono/bi/tri-exponential)

药理学响应与剂量 – 效应Pharmacology & dose-response

激动剂（agonist）的浓度 – 响应曲线，EC₅₀ 计算Agonist concentration-response curve, EC₅₀ determination
拮抗剂（antagonist / blocker）的 IC₅₀ 计算Antagonist/blocker IC₅₀ determination
Hill 系数（n_H）反映协同性Hill coefficient (n_H) for cooperativity
多靶点 / 多化合物并行筛选Multi-target / multi-compound parallel screening

标准服务流程Standard workflow

五步完成从目的基因到电生理数据Five steps from target gene to electrophysiology data

客户提供基因序列、质粒或 cRNAClient provides gene / plasmid / cRNA

载体构建与体外转录 cRNAVector construction & cRNA in-vitro transcription

卵母细胞显微注射（50 nL / 个）Oocyte microinjection (50 nL/cell)

ND96 浴中孵育 1–4 天（18–22℃）Incubation in ND96 bath for 1–4 days (18–22 °C)

TEVC 记录、漏电流校正、曲线拟合、IC₅₀/EC₅₀ 报告TEVC recording, leak subtraction, curve fitting, IC₅₀/EC₅₀ report

参数Parameter	规格Specification
电极类型Electrode type	硼硅酸玻璃微电极，充填 3 M KCl，阻抗 0.5–2 MΩBorosilicate glass microelectrodes, filled with 3 M KCl, resistance 0.5–2 MΩ
标准浴槽液Standard bath	ND96（NaCl 96 mM / KCl 2 mM / CaCl₂ 1.8 mM / MgCl₂ 1 mM / HEPES 5 mM，pH 7.4）
电压钳制范围Voltage clamp range	−150 mV ～ +60 mV（可定制）
采样频率Sampling rate	1–10 kHz（按通道动力学选择）
客户提供Client provides	基因序列 / 质粒 / cRNA；待测化合物（如需）Gene sequence / plasmid / cRNA; test compounds (if required)
交付物Deliverables	完整技术报告 + 原始数据 + 分析图表（I-V / G-V / 剂量 – 效应曲线）Full technical report + raw data + analysis figures (I-V / G-V / dose-response)

精准鉴定你的离子通道Precisely characterize your ion channel

提供目的基因与实验方案，我们完成全流程 TEVC 检测并出具专业报告。Submit your gene and experimental goals — we handle the full TEVC workflow and deliver a professional report.

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