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

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 quiesce...

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Autores principales: Jayant, Krishna, Singhai, Amit, Cao, Yingqiu, Phelps, Joshua B., Lindau, Manfred, Holowka, David A., Baird, Barbara A., Kan, Edwin C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4685269/
https://www.ncbi.nlm.nih.gov/pubmed/26686301
http://dx.doi.org/10.1038/srep18477
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author Jayant, Krishna
Singhai, Amit
Cao, Yingqiu
Phelps, Joshua B.
Lindau, Manfred
Holowka, David A.
Baird, Barbara A.
Kan, Edwin C.
author_facet Jayant, Krishna
Singhai, Amit
Cao, Yingqiu
Phelps, Joshua B.
Lindau, Manfred
Holowka, David A.
Baird, Barbara A.
Kan, Edwin C.
author_sort Jayant, Krishna
collection PubMed
description 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 ([Image: see text]) 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 [Image: see text] fluctuations with a rapid rise and slow decay. Chromaffin cells stimulated with high KCl showed both slow [Image: see text] 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.
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spelling pubmed-46852692015-12-30 Non-Faradaic Electrochemical Detection of Exocytosis from Mast and Chromaffin Cells Using Floating-Gate MOS Transistors Jayant, Krishna Singhai, Amit Cao, Yingqiu Phelps, Joshua B. Lindau, Manfred Holowka, David A. Baird, Barbara A. Kan, Edwin C. Sci Rep Article 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 ([Image: see text]) 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 [Image: see text] fluctuations with a rapid rise and slow decay. Chromaffin cells stimulated with high KCl showed both slow [Image: see text] 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. Nature Publishing Group 2015-12-21 /pmc/articles/PMC4685269/ /pubmed/26686301 http://dx.doi.org/10.1038/srep18477 Text en Copyright © 2015, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Jayant, Krishna
Singhai, Amit
Cao, Yingqiu
Phelps, Joshua B.
Lindau, Manfred
Holowka, David A.
Baird, Barbara A.
Kan, Edwin C.
Non-Faradaic Electrochemical Detection of Exocytosis from Mast and Chromaffin Cells Using Floating-Gate MOS Transistors
title Non-Faradaic Electrochemical Detection of Exocytosis from Mast and Chromaffin Cells Using Floating-Gate MOS Transistors
title_full Non-Faradaic Electrochemical Detection of Exocytosis from Mast and Chromaffin Cells Using Floating-Gate MOS Transistors
title_fullStr Non-Faradaic Electrochemical Detection of Exocytosis from Mast and Chromaffin Cells Using Floating-Gate MOS Transistors
title_full_unstemmed Non-Faradaic Electrochemical Detection of Exocytosis from Mast and Chromaffin Cells Using Floating-Gate MOS Transistors
title_short Non-Faradaic Electrochemical Detection of Exocytosis from Mast and Chromaffin Cells Using Floating-Gate MOS Transistors
title_sort non-faradaic electrochemical detection of exocytosis from mast and chromaffin cells using floating-gate mos transistors
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4685269/
https://www.ncbi.nlm.nih.gov/pubmed/26686301
http://dx.doi.org/10.1038/srep18477
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