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53 Chapter 4 Patch clamp techniques for single channel and whole-cell recording DAVID OGDEN and PETER STANFIELD 1. Introduction The patch clamptechnique.

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Axon Instruments consumables and accessories to satisfy all your patch-clamp needs. A big part of this success has been the ability to provide a comprehensive range.

The HS-2 and HS-2A headstages are used with Axoclamp 2 and GeneClamp amplifiers. They are all unity-gain voltage recording headstages but they come in a variety of different current-passing gains for applications as diverse as extracellular recording, bath-potential recording, ion-sensitive recording, ionophoresis and intracellular recording from small or large cells. For ultra-high impedance electrodes, special circuitry inside the headstage prevents any DC current from leaking into the input through the capacitance neutralization circuit.

HS-2-x0.0001MU

HS-2-x0.01MU

HS-2A-x0.1LU

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HS-2A-x1LU

HS-2A-x1MGU

HS-2A-x10MGU

HS-2A-x100MGU

VG-2A-x100 virtual ground

M versions have extended capacitance neutralization range. G versions have grounded cases. HS-2 A series headstages have an extended operating voltage range 150V.

The benchmark for single-channel recordings and other applications where an excellent signal-to-noise ratio is key.

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Widely accepted as the gold-standard for ultra low-noise patch clamp recordings, the Axopatch 200B Capacitor Feedback Patch Clamp Amplifier is the premier microelectrode amplifier for ultra-low noise patch clamp recordings. Incorporates capacitor-feedback technology and active headstage cooling for single-channel recording, while resistive-feedback circuitry is used for two different ranges of whole-cell recording.

In current-clamp mode, offers two different speeds to optimize membrane potential recording, and allows correction for voltage errors due to pipette resistance. This makes the Axopatch 200B System the amplifier of choice for a scientist who performs basic research in the ion channel field and is the reason why researchers purchased thousands of these amplifiers from Molecular Devices over the past decade.

Actively cooled capacitor-feedback headstage reduces thermal noise for signal-to-noise ratios close to the theoretical limits of physics

Integrating headstage mode for measuring sub-picoamp currents to enable high-fidelity single-channel recordings from cell membranes and artificial bilayers

Resistive headstage mode for larger currents and noise levels in whole-cell recordings

Slim headstage design for easier access to the preparation on a crowded microscope stage

Series resistance compensation: Prediction helps achieve the desired command level faster by transiently supercharging the membrane potential

Series resistance compensation: Correction improves the bandwidth of the recording by eliminating the error introduced by the voltage drop across the series resistance

Dual-speed current clamp optimizes speed and stability through better matching of the clamp algorithm to experimental parameters

Integrated seal test in voltage- and current-clamp mode for easy monitoring of membrane and cell health parameters

Dual command potentials add flexibility by processing input signals from two different sources

Selectable holding command input with two internal range settings and an external input enable convenient control of the output waveform

Pipette capacitance compensation neutralizes the contribution of the glass pipette and the pipette holder to the capacitance of the circuitry

Cell capacitance compensation corrects for the capacitance of the cell membrane in whole-cell recordings

Selectable output gain with 10 settings to scale the output signal to the desired level

Leak subtraction for correction of leak currents without software-based algorithms

Sophisticated 4-pole Bessel low-pass filter acts as anti-aliasing filter and can be used to pre-condition the output signal

Configurable panel meter displays user-selectable parameters in real time for quick monitoring of the experiment

Zap facilitates patch rupturing by applying a voltage pulse for a user-defined duration

Two included pipette holders of different barrel lengths accept glass pipettes in most standard dimensions

User guide written by scientific consultants, with the assistance of Axon Instruments staff, for straightforward start-up and as an in-depth reference

Basic Ion Channel Research

Ion channels are pore proteins that are involved in a large variety of physiological processes. Basic research in the ion channel field furthers the understanding of ion channel function as well as their roles within cellular pathways and processes. Single-channel research helps with elucidating the mechanisms underlying ion channel function on a single-molecule level. Whole-cell and perforated patch clamp recordings are typically used to investigate the action of modulators on entire populations of ion channels in the membrane of a cell. Learn more.

Cell Pathway Analysis

Ion channels are involved in many cell pathways and understanding the function of ion channels in response to changes in membrane potential or the presence or absence of other molecules is important in order to understand exactly how ion channels participate in normal and abnormal biological processes such as cell differentiation and migration, disease states, neuronal communications, etc. Learn more.

Disease Research

Ion channels play a role in many diseases, including hypertension, cardiac arrhythmias, gastrointestinal, immune and neuromuscular disorders, pathological pain and cancer. By understanding the exact role that ion channels play in a particular disease, researchers might be able to find a way to affect the ion channel in such a way as to alter the course of the disease. Learn more.

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Safety Assessment

A critical part of drug discovery involves safety assessments, such as hERG testing, in order to eliminate any compounds with potential safety concerns. Conventional patch clamp has played an important role in collecting evidence for the absence of side effects before submission of a drug candidate to regulatory authorities. Even if automated electrophysiology has shifted safety testing to earlier stages in the drug discovery process, follow-up with conventional patch clamp is an essential step in the workflow. Learn more.

In combination with a Digidata 1550 Data Acquisition System and pCLAMP Software, the Axopatch 200B Amplifier forms the foundation of a patch clamp setup like those used in many electrophysiology labs around the world. Telegraphing between the software and the amplifier ensure correct scaling and logging of experimental parameters. The Clampex Data Acquisition Module, which is part of the pCLAMP Software, makes optimal use of the many advanced features of the Axopatch 200B Amplifier.

General Specifications

General Specifications

Dimensions in.

3.5 H x 19 W x 12.5 D

Dimensions cm

8.9 H x 48.3 W x 31.7 D

Weight

11.5 5.1 kg

Headstage in.

0.75 H x 0.70 W x 4.2 D

Headstage cm

1.8 H x 1.9 W x 10.5 D

Mounting plate in.

0.25 H x 2.0 W x 2.5 D

Mounting plate cm

0.6 H x 5.0 W x 6.2 D

Communications

Analog and digital BNC

Rack use

Standard 19 rack-mount 2U with handles

Benchtop use

Bayonet feet

Power

85-264 VAC 110-340 VDC

50-60 Hz, 30 W max

Fuse

0.5 A slow 15 x 20 mm

Line filter

RFI filter included

Line cord

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Shielded line cord provided

Safety

CE marking Conformité Européenne

Detail Specifications

Configurations

CV 203BU Headstage

Construction:

All critical components are in a sealed hybrid and cooled with a solid state cooling element

Configuration:

High-speed, low-noise current to-voltage converter

Cooling:

Input circuitry -15 C typical. Headstage cooling should be kept on at all times to ensure proper calibration of offset voltages

Gain b :

1 mV/pA b 1 Patch or Whole-Cell modes

0.1 mV/pA b 0.1 Whole-Cell mode

Feedback Element:

Patch

1pF

Whole Cell

β 1, 500 MΩ in parallel with 1 pF

β 0.1, 50 MΩ in parallel with 1 pF

Tuning Whole Cell mode only : Tuning circuit to idealize response of the feedback resistor is contained in the main instrument. Tuning is automatically bypassed when the capacitive feedback is selected.

Pipette-capacitance-compensation injection capacitor: 1 pF

Whole-Cell-Capacitance-Compensation Ranges:

none

β 1 : 0.3-100 pF

β 0.1 : 3-1000 pF

Case: Connected to ground. Case jack mates to 1 mm plugs.

Bandwidth: Test signal applied via Speed Test input

Internal:

Patch mode : Internal: 140 kHz

70 kHz whole-cell mode

External:

100 kHz limited by Bessel filter

Capacitative load stability: 1000 pF, 0 Ω in series

Maximum instrument noise: Measured with minimal external noise sources i.e. radiated line-frequency noise, mechanical vibration 8-pole Bessel filter.

Max. Instrument Noise: With 1-HL-U Holder

Line Frequency and Harmonic

β 1

0.005 pAp-p

β 0.1

0.1 - 100 Hz

0.030 pAp-p

0.50 pAp-p

1.6 pAp-p

0.1 - 1 kHz

0.015 pArms

0.50 pArms

0.75 pArms

0.1 - 5 kHz

0.060 pArms

0.65 pArms

1.65 pArms

0.1 - 10 kHz

0.130 pArms

1.10 pArms

3.0 pArms

Max. Instrument Noise: Without Holder

0.145 pArms

Headstages

Description

Details

Part Number

Axoclamp 900A Headstage HS-9A X0.1U

x0.1 headstage

1-2950-0359

Axoclamp 900A Headstage HS-9A X1U

x1 headstage

1-2950-0360

Axoclamp 900A Headstage HS-9A X10U

x10 headstage

1-2950-0361

Axoclamp 900A Headstage VG-9A X10U

x10 virtual ground headstage

1-2950-0362

Axoclamp 900A Headstage VG-9A X100U

x100 virtual ground headstage

1-2950-0363

MultiClamp 700B Headstage CV-7B

Patch-clamp headstage

1-CV-7B

MultiClamp 700B Headstage CV-7B/BL

Bilayer headstage

1-CV-7B/BL

MultiClamp 700B Headstage CV-7B/EC

Electrochemistry headstage

1-CV-7B/EC

Electrode holders, adapters, and holder components

Electrode Holder for U-Type Headstages

Fits glass pipettes with outer diameter of 1.0 - 1.7 mm

1-HL-U

Electrode Holder Replacement Caps

Set of 2 polycarbonate caps for HL-U holders

1-HL-CAP

Cone Washers 1.1mm ID

Set of 10 orange cone washers for HL-U holders, fit glass with outer diameter of 1.0 - 1.1 mm

1-HLC-11

Cone Washers 1.3mm ID

Set of 10 orange cone washers for HL-U holders, fit glass with outer diameter of 1.1 - 1.3 mm

1-HLC-13

Cone Washers 1.5mm ID

Set of 10 orange cone washers for HL-U holders, fit glass with outer diameter of 1.3 - 1.5 mm

1-HLC-15

Cone Washers 1.7mm ID

Set of 10 orange cone washers for HL-U holders, fit glass with outer diameter of 1.5 - 1.7 mm

1-HLC-17

Pins 1mm for HL-U Holders

Set of 3 brass pins for HL-U holders, 1mm

1-HLP-U

2mm Plugs with Solder Cups

Set of 5 general purpose gold plugs, 2mm, with solder cups

1-HLP-0

Silver Wire

Set of 5 Ag wires, 0.25mm diameter, 50mm long

1-HLA-005

Silicone Tubing for Silver Wire

1mm ID x 70mm Long Silicone Tubing

1-HLT-70

Silver/Silver Chloride Pellet Assemblies

Set of 3 Ag/AgCl pellet assemblies

1-HLA-003

Adapter For BNC Holders To U-Type Headstages

Connects BNC holders to CV and HS headstages with threaded collets U-type

1-HLB-U

Right-Angle Adapter for HL-U Electrode Holders

Fits CV and HS headstages with threaded collets U-type

1-HLR-U

Model Cells

Model Cell for Oocytes

Axoclamp/ GeneClamp model cell for oocytes. Connects to U-type HS series headstages

1-MCO-2U

Model Cell for TEVC / DSEVC

Axoclamp/ GeneClamp model cell for two-electrode voltage clamp/ discontinuous single-electrode voltage-clamp conditions. Connects to U-type HS series headstages

1-CLAMP-1U

Model Cell for Whole Cell/ Single Channels

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Axopatch/ GeneClamp/ MultiClamp model cell for whole-cell / single-channel patch-clamp conditions. Connects to U-type CV series headstages

1-PATCH-1U

Model Cell for Bilayers

Axopatch/ GeneClamp/ MultiClamp model cell for bilayer conditions. Connects to U-type CV series headstages

1-MCB-1U

Cables

Cable To Connect Axoclamp 2 Headstages To Axoclamp 900A Amplifier

Allows Axoclamp 2 headstages HS-2, VG-2 to be used on Axoclamp 900A Amplifiers

1-2100-0934

Miscellaneous

SoftPanel Amplifier Control Unit

Provides physical knob and button control for computer-controlled Axoclamp 900-series and MultiClamp 700-series amplifiers. Requires a USB connection.

1-SOFTPANEL USB

Remote Buzz For Axoclamp 900A

Hand-held buzz duration control for Axoclamp 900A Amplifier 1-50ms

1-2950-0366

Set of 3 Ag/AgCl pellet assembly and Ag wire

Number of Citations : 26,200

Latest Citations:

For a complete list of citations, please click here.

Ion Selectivity of Lysenin Channels

J May, S Bryant, M Johnson, R Good - 2015 - scholarworks.boisestate.edu

The solutions bathing the membrane are wired to an Axopatch 200B electrophysiology

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amplifier through Ag/AgCl electrodes. The solutions bathing the membrane are wired to an

Axopatch 200B electrophysiology amplifier through Ag/AgCl electrodes.

Chemical activation of the mechanotransduction channel Piezo1

R Syeda, J Xu, AE Dubin, B Coste, J Mathur, T Huynh - Elife, 2015 - elifesciences.org

Stretch-activated currents were recorded using 244 Axopatch 200B amplifier Molecular Devices

Axopatch 200B. ratio. Electrode carrying the proteoliposome droplet was connected to the working

end of the 276 amplifier head-stage Molecular Devices Axopatch 200B.

Murine and human CFTR exhibit different sensitivities to CFTR potentiators

G Cui, NA McCarty - Journal of Physiology-Lung Cellular and , 2015 - Am Physiological Soc

and 127.6 U/ml PKA pH 7.5. CFTR currents were measured with an Axopatch 200B

amplifier Molecular Devices; Sunnyvale, CA, and were recorded at 10 kHz to DAT

tape. For subsequent analysis, records were played back

Functional characterization of rare variants implicated in susceptibility to lone atrial fibrillation

K Hayashi, T Konno, H Tada, S Tani, L Liu - Circulation: , 2015 - Am Heart Assoc

Potassium or sodium currents were studied using the whole-cell patch clamp technique with

an amplifier, Axopatch-200B Molecular Devices, Sunnyvale, CA, Two-tailed patch clamp

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technique with an amplifier, Axopatch-200B Molecular Devices, Suununnynyn vale, CA CA,

as of November 13, 2015. Source: Google Scholar. Search results include Axopatch 200B and Axopatch Axon Instruments.

Widely accepted as the gold-standard for ultra low-noise patch clamp recordings, the Axopatch 200B Capacitor Feedback Patch Clamp Amplifier is the premier.

The BVC-700A is a microelectrode preamplifier with both Bridge Current Clamp and Continuous Voltage Clamp Capability. The BVC-700A was designed to serve a wide.

Feb 25, 2014 In this second part of the unscripted video, I give you a very simplistic overview of what equipment you might need or encounter when setting up a single.