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Non-Faradaic Electrochemical Detection of Exocytosis from Mast and Chromaffin Cells Using Floating-Gate MOS Transistors

机译:使用浮动门MOS晶体管的非法拉第电化学检测肥大细胞和嗜铬细胞的胞吐作用

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摘要

We present non-faradaic electrochemical recordings of exocytosis from populations of mast and chromaffin cells using chemoreceptive neuron MOS (CνMOS) transistors. In comparison to previous cell-FET-biosensors, the CνMOS features control (CG), sensing (SG) and floating gates (FG), allows the quiescent point to be independently controlled, is CMOS compatible and physically isolates the transistor channel from the electrolyte for stable long-term recordings. We measured exocytosis from RBL-2H3 mast cells sensitized by IgE (bound to high-affinity surface receptors FcεRI) and stimulated using the antigen DNP-BSA. Quasi-static I-V measurements reflected a slow shift in surface potential () which was dependent on extracellular calcium ([Ca]o) and buffer strength, which suggests sensitivity to protons released during exocytosis. Fluorescent imaging of dextran-labeled vesicle release showed evidence of a similar time course, while un-sensitized cells showed no response to stimulation. Transient recordings revealed fluctuations with a rapid rise and slow decay. Chromaffin cells stimulated with high KCl showed both slow shifts and extracellular action potentials exhibiting biphasic and inverted capacitive waveforms, indicative of varying ion-channel distributions across the cell-transistor junction. Our approach presents a facile method to simultaneously monitor exocytosis and ion channel activity with high temporal sensitivity without the need for redox chemistry.
机译:我们目前使用化学感受性神经元MOS(CνMOS)晶体管从肥大和嗜铬细胞的种群中的胞吐作用的非法拉第电化学记录。与以前的电池FET生物传感器相比,CνMOS具有控制(CG),感应(SG)和浮栅(FG)的功能,允许对静态点进行独立控制,与CMOS兼容并将晶体管通道与电解质物理隔离稳定的长期录音。我们测量了由IgE致敏的RBL-2H3肥大细胞(与高亲和力表面受体FcεRI结合)并使用抗原DNP-BSA刺激的胞吐作用。准静态的I-V测量反映了表面电势()的缓慢变化,这取决于细胞外钙(Ca)和缓冲液的强度,这表明对胞吐过程中释放的质子敏感。右旋糖酐标记的囊泡释放的荧光成像显示了类似的时程证据,而未敏化的细胞对刺激没有反应。瞬态记录显示波动迅速而缓慢衰减。用高KCl刺激的嗜铬细胞既显示缓慢移动,又显示出双相和反向电容波形的细胞外动作电位,表明跨细胞-晶体管连接的离子通道分布变化。我们的方法提出了一种简便的方法,可以以高时间敏感性同时监测胞吐作用和离子通道活性,而无需氧化还原化学。

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