Non-Faradaic Electrochemical Detection of Exocytosis from Mast and Chromaffin Cells Using Floating-Gate MOS Transistors.
Non-Faradaic Electrochemical Detection of Exocytosis from Mast and Chromaffin Cells Using Floating-Gate MOS Transistors.
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使用浮栅 MOS 晶体管对肥大细胞和嗜铬细胞胞吐作用进行非法拉第电化学检测。
DOI：
10.1038/srep18477
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发表时间：
2015
影响因子：
4.6
通讯作者：
Kan,EdwinC
中科院分区：
文献类型：
--
作者：
Jayant,Krishna;Singhai,Amit;Cao,Yingqiu;Phelps,JoshuaB;Lindau,Manfred;Holowka,DavidA;Baird,BarbaraA;Kan,EdwinC
我们提出了非法拉第电化学记录的肥大细胞和嗜铬细胞的胞吐使用化学感受神经元MOS（CV MOS）晶体管。与以前的细胞FET生物传感器相比，C V MOS具有控制（CG），感测（SG）和浮栅（FG），允许独立控制静态点，是CMOS兼容的，并将晶体管沟道与电解质物理隔离，以实现稳定的长期记录。我们测量了由IgE（结合高亲和力表面受体FcεRI）致敏并使用抗原DNP-BSA刺激的RBL-2 H3肥大细胞的胞吐作用。准静态IV测量反映了表面电位（）的缓慢变化，其取决于细胞外钙（[Ca] o）和缓冲强度，这表明对胞吐期间释放的质子敏感。葡聚糖标记的囊泡释放的荧光成像显示出类似的时间过程的证据，而未致敏的细胞显示出对刺激没有反应。瞬态记录揭示了快速上升和缓慢衰减的波动。高氯化钾刺激的嗜铬细胞表现出缓慢的位移和细胞外动作电位表现出双相和反向电容波形，表明不同的离子通道分布在整个细胞晶体管结。我们的方法提出了一种简便的方法，同时监测胞吐作用和离子通道的活动与高时间灵敏度，而不需要氧化还原化学。
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 IV 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.