Achieving maximal speed of solution exchange for patch clamp experiments

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Background: Resolving the kinetics of agonist binding events separately from the subsequent channel gating processes requires the ability of applying and removing the agonist before channel gating occurs. No reported system has yet achieved pulses shorter than 100 μs, necessary to study nicotinic AC...

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Autores principales: Auzmendi, J., Fernández Do Porto, D., Pallavicini, C., Moffatt, L.
Formato: Artículo publishedVersion
Lenguaje:Inglés
Publicado: 2012
Materias:
agonist
analytical parameters
article
channel gating
electric potential
flow rate
measurement
oscillation
patch clamp
piezoelectricity
pipette
process optimization
solution exchange system
Electricity
Movement
Patch-Clamp Techniques
Rheology
Solutions
Time Factors
Acceso en línea:http://hdl.handle.net/20.500.12110/paper_19326203_v7_n8_p_Auzmendi
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Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires
id paperaa:paper_19326203_v7_n8_p_Auzmendi
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spelling paperaa:paper_19326203_v7_n8_p_Auzmendi2023-06-12T16:51:42Z Achieving maximal speed of solution exchange for patch clamp experiments PLoS ONE 2012;7(8) Auzmendi, J. Fernández Do Porto, D. Pallavicini, C. Moffatt, L. agonist analytical parameters article channel gating electric potential flow rate measurement oscillation patch clamp piezoelectricity pipette process optimization solution exchange system Electricity Movement Patch-Clamp Techniques Rheology Solutions Time Factors Background: Resolving the kinetics of agonist binding events separately from the subsequent channel gating processes requires the ability of applying and removing the agonist before channel gating occurs. No reported system has yet achieved pulses shorter than 100 μs, necessary to study nicotinic ACh receptor or AMPA receptor activation. Methodology/Principal Findings: Solution exchange systems deliver short agonist pulses by moving a sharp interface between a control and an experimental solution across a channel preparation. We achieved shorter pulses by means of an exchange system that combines a faster flow velocity, narrower partition between the two streams, and increased velocity and bandwidth of the movement of the interface. The measured response of the entire system was fed back to optimize the voltage signal applied to the piezoelectric actuator overcoming the spurious oscillations arising from the mechanical resonances when a high bandwidth driving function was applied. Optimization was accomplished by analyzing the transfer function of the solution exchange system. When driven by optimized command pulses the enhanced system provided pulses lasting 26 ± 1 μs and exchanging 93 ± 1% of the solution, as measured in the open tip of a patch pipette. Conclusions/Significance: Pulses of this duration open the experimental study of the molecular events that occur between the agonist binding and the opening of the channel. © 2012 Auzmendi et al. Fil:Auzmendi, J. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Fernández Do Porto, D. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Moffatt, L. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2012 info:eu-repo/semantics/article info:ar-repo/semantics/artículo info:eu-repo/semantics/publishedVersion application/pdf eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_19326203_v7_n8_p_Auzmendi
institution Universidad de Buenos Aires
institution_str I-28
repository_str R-134
collection Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
language Inglés
orig_language_str_mv eng
topic agonist
analytical parameters
article
channel gating
electric potential
flow rate
measurement
oscillation
patch clamp
piezoelectricity
pipette
process optimization
solution exchange system
Electricity
Movement
Patch-Clamp Techniques
Rheology
Solutions
Time Factors
spellingShingle agonist
analytical parameters
article
channel gating
electric potential
flow rate
measurement
oscillation
patch clamp
piezoelectricity
pipette
process optimization
solution exchange system
Electricity
Movement
Patch-Clamp Techniques
Rheology
Solutions
Time Factors
Auzmendi, J.
Fernández Do Porto, D.
Pallavicini, C.
Moffatt, L.
Achieving maximal speed of solution exchange for patch clamp experiments
topic_facet agonist
analytical parameters
article
channel gating
electric potential
flow rate
measurement
oscillation
patch clamp
piezoelectricity
pipette
process optimization
solution exchange system
Electricity
Movement
Patch-Clamp Techniques
Rheology
Solutions
Time Factors
description Background: Resolving the kinetics of agonist binding events separately from the subsequent channel gating processes requires the ability of applying and removing the agonist before channel gating occurs. No reported system has yet achieved pulses shorter than 100 μs, necessary to study nicotinic ACh receptor or AMPA receptor activation. Methodology/Principal Findings: Solution exchange systems deliver short agonist pulses by moving a sharp interface between a control and an experimental solution across a channel preparation. We achieved shorter pulses by means of an exchange system that combines a faster flow velocity, narrower partition between the two streams, and increased velocity and bandwidth of the movement of the interface. The measured response of the entire system was fed back to optimize the voltage signal applied to the piezoelectric actuator overcoming the spurious oscillations arising from the mechanical resonances when a high bandwidth driving function was applied. Optimization was accomplished by analyzing the transfer function of the solution exchange system. When driven by optimized command pulses the enhanced system provided pulses lasting 26 ± 1 μs and exchanging 93 ± 1% of the solution, as measured in the open tip of a patch pipette. Conclusions/Significance: Pulses of this duration open the experimental study of the molecular events that occur between the agonist binding and the opening of the channel. © 2012 Auzmendi et al.
format Artículo
Artículo
publishedVersion
author Auzmendi, J.
Fernández Do Porto, D.
Pallavicini, C.
Moffatt, L.
author_facet Auzmendi, J.
Fernández Do Porto, D.
Pallavicini, C.
Moffatt, L.
author_sort Auzmendi, J.
title Achieving maximal speed of solution exchange for patch clamp experiments
title_short Achieving maximal speed of solution exchange for patch clamp experiments
title_full Achieving maximal speed of solution exchange for patch clamp experiments
title_fullStr Achieving maximal speed of solution exchange for patch clamp experiments
title_full_unstemmed Achieving maximal speed of solution exchange for patch clamp experiments
title_sort achieving maximal speed of solution exchange for patch clamp experiments
publishDate 2012
url http://hdl.handle.net/20.500.12110/paper_19326203_v7_n8_p_Auzmendi
work_keys_str_mv AT auzmendij achievingmaximalspeedofsolutionexchangeforpatchclampexperiments
AT fernandezdoportod achievingmaximalspeedofsolutionexchangeforpatchclampexperiments
AT pallavicinic achievingmaximalspeedofsolutionexchangeforpatchclampexperiments
AT moffattl achievingmaximalspeedofsolutionexchangeforpatchclampexperiments
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