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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.
Autopatcher.org In Vivo Robotics, for Automatic Recording of Neurons in the Live Brain.
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
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:
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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
Forefront Tmg 2010 Keygen Generator Acids
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.