Patch Clamp Recording in Vivo

2015 ◽  
Author(s):  
David Ferster
Keyword(s):  
Patch Clamp ◽  
Brain Slices ◽  
Mammalian Brain ◽  
Intracellular Recordings ◽  
Intracellular Membrane ◽  
Patch Clamp Recording ◽  
Intact Organism ◽  
Technological Developments ◽  
History Of

Patch clamp recording in vivo allows an investigator to study intracellular membrane potentials in an intact organism (as opposed to cells in culture or acute brain slices). This technique is a reliable method of obtaining high-quality intracellular recordings from neurons, regardless of their size, in several parts of the mammalian brain. This chapter will describe the principles and practice of performing patch clamp experiments in vivo, beginning with a brief history of the technological developments that have made this technique possible.

Patch Clamp Recording in Brain Slices

2015 ◽  
Author(s):  
Luke Campagnola ◽  
Paul Manis
Keyword(s):  
Patch Clamp ◽  
Brain Slices ◽  
Neuronal Membrane ◽  
Electrical Noise ◽  
Patch Clamp Recording ◽  
Experimental Conditions ◽  
Neural Substrate ◽  
Hardware Configuration ◽  
Technical Issues

Patch clamp recording in brain slices allows unparalleled access to neuronal membrane signals in a system that approximates the in-vivo neural substrate while affording greater control of experimental conditions. In this chapter we discuss the theory, methodology, and practical considerations of such experiments including the initial setup, techniques for preparing and handling viable brain slices, and patching and recording signals. A number of practical and technical issues faced by electrophysiologists are also considered, including maintaining slice viability, visualizing and identifying healthy cells, acquiring reliable patch seals, amplifier compensation features, hardware configuration, sources of electrical noise and table vibration, as well as basic data analysis issues and some troubleshooting tips.

scholarly journals Automated in vivo patch-clamp evaluation of extracellular multielectrode array spike recording capability

2018 ◽  
Vol 120 (5) ◽  
pp. 2182-2200 ◽  
Author(s):  
Brian D. Allen ◽  
Caroline Moore-Kochlacs ◽  
Jacob G. Bernstein ◽  
Justin P. Kinney ◽  
Jorg Scholvin ◽  
...  
Keyword(s):  
Patch Clamp ◽  
Brain Regions ◽  
Automated System ◽  
Mammalian Brain ◽  
Multielectrode Array ◽  
Spike Sorting ◽  
Multielectrode Arrays ◽  
Patch Clamp Recording ◽  
Data Set

Much innovation is currently aimed at improving the number, density, and geometry of electrodes on extracellular multielectrode arrays for in vivo recording of neural activity in the mammalian brain. To choose a multielectrode array configuration for a given neuroscience purpose, or to reveal design principles of future multielectrode arrays, it would be useful to have a systematic way of evaluating the spike recording capability of such arrays. We describe an automated system that performs robotic patch-clamp recording of a neuron being simultaneously recorded via an extracellular multielectrode array. By recording a patch-clamp data set from a neuron while acquiring extracellular recordings from the same neuron, we can evaluate how well the extracellular multielectrode array captures the spiking information from that neuron. To demonstrate the utility of our system, we show that it can provide data from the mammalian cortex to evaluate how the spike sorting performance of a close-packed extracellular multielectrode array is affected by bursting, which alters the shape and amplitude of spikes in a train. We also introduce an algorithmic framework to help evaluate how the number of electrodes in a multielectrode array affects spike sorting, examining how adding more electrodes yields data that can be spike sorted more easily. Our automated methodology may thus help with the evaluation of new electrode designs and configurations, providing empirical guidance on the kinds of electrodes that will be optimal for different brain regions, cell types, and species, for improving the accuracy of spike sorting. NEW & NOTEWORTHY We present an automated strategy for evaluating the spike recording performance of an extracellular multielectrode array, by enabling simultaneous recording of a neuron with both such an array and with patch clamp. We use our robot and accompanying algorithms to evaluate the performance of multielectrode arrays on supporting spike sorting.

scholarly journals Imaging Subthreshold Voltage Oscillation With Cellular Resolution in the Inferior Olive in vitro

2020 ◽  
Vol 14 ◽  
Author(s):  
Kevin Dorgans ◽  
Bernd Kuhn ◽  
Marylka Yoe Uusisaari
Keyword(s):  
Inferior Olive ◽  
Brain Slices ◽  
Brain Regions ◽  
Mammalian Brain ◽  
Wide Field ◽  
Voltage Imaging ◽  
Subthreshold Oscillations ◽  
Cellular Resolution

Voltage imaging with cellular resolution in mammalian brain slices is still a challenging task. Here, we describe and validate a method for delivery of the voltage-sensitive dye ANNINE-6plus (A6+) into tissue for voltage imaging that results in higher signal-to-noise ratio (SNR) than conventional bath application methods. The not fully dissolved dye was injected into the inferior olive (IO) 0, 1, or 7 days prior to acute slice preparation using stereotactic surgery. We find that the voltage imaging improves after an extended incubation period in vivo in terms of labeled volume, homogeneous neuropil labeling with saliently labeled somata, and SNR. Preparing acute slices 7 days after the dye injection, the SNR is high enough to allow single-trial recording of IO subthreshold oscillations using wide-field (network-level) as well as high-magnification (single-cell level) voltage imaging with a CMOS camera. This method is easily adaptable to other brain regions where genetically-encoded voltage sensors are prohibitively difficult to use and where an ultrafast, pure electrochromic sensor, like A6+, is required. Due to the long-lasting staining demonstrated here, the method can be combined, for example, with deep-brain imaging using implantable GRIN lenses.

In Vivo Patch-Clamp Recording in Awake Head-Fixed Rodents

2017 ◽  
Vol 2017 (4) ◽  
pp. pdb.prot095802 ◽  
Author(s):  
Doyun Lee ◽  
Albert K. Lee
Keyword(s):  
Patch Clamp ◽  
Patch Clamp Recording

scholarly journals Tolerance to Sedative/Hypnotic Actions of GABAergic Drugs Correlates With Tolerance to Potentiation of Extrasynaptic Tonic Currents of Alcohol-Dependent Rats

2009 ◽  
Vol 102 (1) ◽  
pp. 224-233 ◽  
Author(s):  
Jing Liang ◽  
Igor Spigelman ◽  
Richard W. Olsen
Keyword(s):  
Hippocampal Neurons ◽  
Brain Slices ◽  
Brain Regions ◽  
Reasonable Assumption ◽  
Total Charge ◽  
Cross Tolerance ◽  
Patch Clamp Recording ◽  
Anesthetic Drugs ◽  
Drug Actions

Alcohol tolerance resulting from chronic administration is well known to be accompanied by cross-tolerance to sedative/anesthetic drugs, especially those acting on the γ-aminobutyric acid type A receptors (GABAARs). Rats treated with chronic intermittent ethanol (CIE) show decreased function and altered pharmacology of GABAARs in hippocampal neurons, consistent with cell- and location-specific changes in GABAAR subunit composition. We previously observed variably altered sensitivity to GABAergic drugs in vivo and in hippocampal neurons using whole cell patch-clamp recording in brain slices. Here, we examined additional clinical GABAergic drugs to correlate CIE-induced tolerance to potentiation of neuronal GABAAR-mediated currents with tolerance of these agents to sedative/anesthetic effects in vivo. Typical of several drug classes and two cell types, in CA1 pyramidal neurons, the benzodiazepine diazepam doubled the total charge transfer (TCT) of miniature postsynaptic inhibitory currents (mIPSCs), whereas it quadrupled the TCT of tonic currents. CIE treatment altered these responses to variable extent, as it did to loss of righting reflex (LORR) induced by these same drugs: 90–95% tolerance to flurazepam, the neuroactive steroid alphaxalone, and ethanol; 30–40% to pentobarbital, etomidate, and the GABA agonist gaboxadol; and no tolerance to propofol. There was a strong correlation between tolerance in the LORR assay and tolerance to enhancement of tonic currents, but not mIPSCs. The striking correlation suggests that the sedative/anesthetic actions of GABAergic drugs may be mediated primarily via the potentiation of extrasynaptic GABAARs. This requires the reasonable assumption that the same types of GABAARs in other brain regions involved directly in hypnotic drug actions show similar tolerance.

scholarly journals Combined single cell RT-PCR analysis with patch clamp recording in acute brain slices

2000 ◽  
Vol 82 ◽  
pp. 12
Author(s):  
Keisuke Tsuzuki ◽  
Bertrand Lambolez ◽  
Etienne Audinat ◽  
James T. Porter ◽  
Bruno Cauli ◽  
...  
Keyword(s):  
Patch Clamp ◽  
Single Cell ◽  
Brain Slices ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Patch Clamp Recording

Changes in synaptic transmission of substantia gelatinosa neurons in a rat model of lumbar radicular pain revealed by in vivo patch-clamp recording

Pain ◽  
2011 ◽  
Vol 152 (5) ◽  
pp. 1024-1032 ◽  
Author(s):  
Yoshinori Terashima ◽  
Mikito Kawamata ◽  
Tsuneo Takebayashi ◽  
Satoshi Tanaka ◽  
Katsumasa Tanimoto ◽  
...  
Keyword(s):  
Patch Clamp ◽  
Synaptic Transmission ◽  
Rat Model ◽  
Radicular Pain ◽  
Substantia Gelatinosa ◽  
Patch Clamp Recording ◽  
Lumbar Radicular Pain
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