Membrane patch patch clamp techniques. Medical search. Frequent questions

This clamp is critical for testing voltage dependent opening dynamics of ion channels which determine a membrane's resting potentials and action potentials. (wikipedia.org)
The ability of excitable tissue to generate and propagate action potentials depends upon the electrical properties of the cell membrane at rest. (wfsahq.org)

The coupling of the decrease in P o 2 to the exocytotic machinery responsible for the release of neurotransmitters in chemoreceptor cells (i.e., the chemotransduction process) is incompletely understood, but it is well documented that plasma membrane mechanisms are involved. (aspetjournals.org)
While plasma membrane ion channels have been extensively studied, much less is known about the identity and physiology of intracellular channels because they are less accessible to direct electrophysiological characterization. (elifesciences.org)
Methods: Using in situ patch clamp recordings and pharmacological manipulation of astrocytes in CA1 region of the mouse hippocampal brain slices, we investigated transport of D-serine across the plasma membrane. (mendeley.com)

Ion channels are membrane proteins that regulate the concentration of key signaling ions to control a wide range of cellular functions. (elifesciences.org)
With a throughput of up to 10,000 data points per day, the SURFE²R 96SE is a versatile tool for hit validation with the unique feature of accessing proteins residing in their native membrane environment, including organelles. (nanion.de)

Patch-seq is a specialized form of the patch clamp recording technique, the gold standard of single cell electrophysiological studies for its millisecond resolution of cellular electrophysiology, its ability to detect currents of specific ions, and perhaps most importantly for its ability to form a voltage clamp on the cell membrane. (wikipedia.org)
Initially the need for direct contact between the pipette and the cell membrane limited patch clamp's use to cultured cell preparations. (wikipedia.org)
The newly discovered pores of the inner (cell) membrane of bacteria will be studied taking advantage of their ion-conducting properties. (europa.eu)
The Chemical composition of the cytosol and the extracellular fluid, as well as that of the cell membrane itself, are crucial to its electrical properties. (wfsahq.org)

While other modern recording techniques such as multi-electrode array and fluorescent calcium sensors have emerged as alternatives which can record from a great deal more neurons at once, patch clamp offers millisecond temporal resolution and the ability to study specific ionic currents. (wikipedia.org)
Consistent with the binding, functional analysis of pentobarbital-enhanced GABA currents recorded with whole-cell patch clamp demonstrated the β 2 (G219F) subunit mutation eliminated the potentiating effect of the anesthetic. (aspetjournals.org)
We investigated the effect of almitrine on ionic currents of chemoreceptor cells isolated from the carotid body of rat and rabbits by using the whole-cell and inside-out configurations of the patch-clamp technique. (aspetjournals.org)
This effect of almitrine on the Ca 2+ -dependent component of K + currents was investigated further at the single-channel level in excised patches in the inside-out configuration. (aspetjournals.org)
Just like the SURFE 2 R N1, the SURFE 2 R 96SE employs solid supported membrane (SSM)-based electrophysiology to resolve small currents of transporters and pumps but elevates the throughput to the next level by parallelization of 96 measurements. (nanion.de)
It involved placing a tiny glass patch pipette, onto the surface of the astrocytic membrane, to record D-serine transporter currents. (mendeley.com)
1. The extracellular patch clamp method, which first allowed the detection of single channel currents in biological membranes, has been further refined to enable higher current resolution, direct membrane patch potential control, and physical isolation of membrane patches. (diavacs.com)
Using patch-clamp techniques, excitation and secretion in chromaffin cells were studied by measurement of unitary inward currents and of stimulus-evoked increments in membrane capacitance. (mpg.de)
This study therefore aimed to investigate the effects of these agents on membrane currents, elevations of [Ca2+] and contractions evoked by depolarization and/or acetylcholine in airway smooth muscle. (mcmaster.ca)
Patch-clamp, fura-2 fluorimetric and muscle-bath techniques were used to monitor ion currents, [Ca2+] and contractions, respectively, in canine tracheal smooth muscle in the presence and absence of the chromones. (mcmaster.ca)
Exogenous ascorbate also induces a moderate increase in programmed cell death symptoms in intact roots, but it does not activate Ca2+ influx currents in patch-clamped root protoplasts. (deepdyve.com)

Patch-sequencing (patch-seq) is a method designed for tackling specific problems involved in characterizing neurons. (wikipedia.org)
By combining multiple data modalities such as electrophysiology, sequencing and microscopy, Patch-seq allows for neurons to be characterized in multiple ways simultaneously. (wikipedia.org)
Patch clamp can be used to record from adjacent and nearby cells to understand how small cliques of neurons connect to each other and study sub-threshold events. (wikipedia.org)
Membrane properties of fast-spiking interneurons were distinguished from those of medium-sized spiny neurons by their more depolarized resting membrane potential, lower action potential amplitude and shorter half-width, short spike repolarization time and deep spike afterhyperpolarization. (nih.gov)
In conclusion, ventral striatal fast-spiking interneurons can be clearly distinguished from medium-sized spiny neurons by their membrane properties and their firing patterns can be subdivided in bursting and non-bursting modes. (nih.gov)
Using perforated whole-cell patch clamp, we simultaneously monitored all synaptic connections within the networks of three or four neurons. (jneurosci.org)
Genetic engineering techniques allow researchers to tag neurons of interest with fluorescent markers that glow upon neural activation, but capturing those brain areas in action, both as distinct neural circuits and at the resolution of single cells, can be hard. (the-scientist.com)
Perforated patch-clamp analysis of the passive membrane properties of three classes of hippocampal neurons. (janelia.org)
1. Perforated patch-clamp recordings were made from the three major classes of hippocampal neurons in conventional in vitro slices prepared from adult guinea pigs. (janelia.org)
4. It was not possible to measure purely passive membrane properties that were completely independent of membrane potential in any of the three classes of hippocampal neurons. (janelia.org)

Both Erwin Neher and Bert Sakmann, the inventors of the technique, went on to win the 1991 Nobel Prize in Physiology or Medicine for developing a patch clamp and using it to prove the existence of ion channels. (wikipedia.org)

The technique was originally developed using a small glass tube, pipette, which is rapidly heated and stretched to produce a needle like shape with an opening diameter of 3-5 millimeters. (wikipedia.org)
The chance of the pipette tip clamping off target was reduced via application of positive pressure during insertion or use of a cleaning pipette to ensure the main seal was not damaged. (wikipedia.org)
Intriguingly, the replacement of gluconate with ascorbate in the patch-clamp pipette reveals a large ascorbate efflux current, which shows sensitivity to the anion channel blocker, anthracene-9-carboxylic acid (A9C), indicative of the ascorbate release via anion channels. (deepdyve.com)

2016). AA also takes part in the regeneration of α-tocopherol (vitamin E) in photosynthetic membranes and participates in chloroplast metabolism as a cofactor for violaxanthin de-epoxidase, an enzyme involved in xanthophyll cycle-mediated photoprotection (Smirnoff and Wheeler, 2000). (deepdyve.com)

Patch recordings yield an approximate channel density of 7 pS/micron^2 in juvenile rats (yale.edu)

The high resistance seal allows the experimenter to hold the patch of membrane within the seal at a desired voltage for the study of voltage-gated ion channels. (wikipedia.org)
Researchers have successfully recorded current in membrane channels of contracting cardiac cells using the patch clamp technique and an atomic force microscope. (neurosciencenews.com)
In cell-attached patches, single ATP-sensitive K + (K ATP ) channels were activated by bath-applied insulin and subsequently inhibited by wortmannin. (diabetesjournals.org)
High conductance, stretch activated channels have been discovered in both Gram positive and Gram negative bacteria and located in the inner membrane. (europa.eu)
The channels were inhibited again by increasing the concentration of lanthanides, suggesting variation in the mechanical properties of the membrane possessing an extremely high number of binding sites of varying affinity for the trivalents. (europa.eu)
The inner membrane fraction of Escherichia coli, fused into giant liposomes and studied by patch clamp, revealed several other channels which were insensitive to the applied suction. (europa.eu)
Further biochemical characterisation of the channels will be carried out using a traditional isolation approach involving solubilization and reconstitution of protein fractions (the most convenient functional assay being provided by the planar bilayer technique), leading to a study of the molecular biology of the channels. (europa.eu)

Lipid bilayer membranes are among the most ubiquitous structures in the living world, with intricate structural features and a multitude of biological functions. (nature.com)
The lipid bilayer is a critical component of all biological membranes , including cell membranes , and so is absolutely essential for all life on Earth. (wikidoc.org)
The interfacial regions of model phospholipid bilayers have a thickness of 8 to 10 Å, although they can be wider in biological membranes that include lipid molecules whose head groups have complex carbohydrates, found in the gangliosides or lipopolysaccharides [1] . (wikidoc.org)
This is close to the hydrophobic thickness of typical biological membranes measured by small angle X-ray scattering (27-32 Å) [3] . (wikidoc.org)
Develop a biological knowledge base common to contemporary biologists, encompassing microscopy, cell structure, cellular membranes, and nucleic acids biology. (uaeu.ac.ae)

The difference in electrical charge across the membrane is called the Resting Membrane Potential . (wfsahq.org)

We combined this with whole cell patch-clamp recording techniques to examine the physiological properties of CCK-containing interneurons of the rat BLA. (nih.gov)
we used hierarchical cluster and discriminant function analysis to demonstrate that CCK interneurons can be segregated into three distinct subtypes (I, II, III) based on their passive and active membrane properties. (nih.gov)
In vitro patch-clamp recordings were made to study the membrane properties and synaptic connectivity of fast-spiking interneurons in rat ventral striatum. (nih.gov)

A lipid bilayer or bilayer lipid membrane (BLM) is a membrane or zone of a membrane composed of lipid molecules (usually phospholipids ). (wikidoc.org)
They concluded, correctly, that the membrane is two lipid molecules thick and proposed it is made of a bilayer. (wikidoc.org)
Support for the existence of a lipid bilayer in cell membranes came with the discovery by Alec Bangham in 1965 that phospholipids , when introduced into an aqueous environment, spontaneously form liposomes . (wikidoc.org)
The properties of the bilayer are influenced by a variety of factors, including the lipid composition, temperature and membrane pressure. (wikidoc.org)

The Swarm delivers variable intensity blue-light optogenetic stimulation to enable membrane depolarization and red or lime-light excitation to enable fluorescence detection of the resulting changes in membrane potential or calcium levels, respectively. (frontiersin.org)

ECT constructs made by repopulating cell-free scaffolds with suitable cells are usually referred to as cardiac patches. (encyclopedia.pub)

The bilayer is composed of two layers of lipids arranged so that their hydrocarbon tails face one another to form an oily core held together by Van der Waals interactions, while their charged heads face the aqueous solutions on either side of the membrane. (wikidoc.org)

In addition, the presence of the flat cholesterol molecule in the membrane tends to straighten out the hydrocarbon chains, causing two main effects: the membrane becomes even less permeable to small molecules and the thickness of the hydrocarbon region is increased. (wikidoc.org)

used a technique called patch-clamp to study the movement of ions across the membrane of melanosomes from skin and eye cells. (elifesciences.org)
To monitor brain activity in mice and other animals, neuroscientists often rely on a well-established technique called patch clamping, which can ignite career-questioning frustration, as electrical noise spoils seemingly good data and cells begin to die after only a few seconds of recording. (the-scientist.com)
Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. (yale.edu)
Using the perforated patch clamp technique, we found that insulin (1-600 nmol/l) but not IGF-1 (100 nmol/l) reversibly hyperpolarized single mouse β-cells and inhibited their electrical activity. (diabetesjournals.org)
Human Embryonic Kidney Hek 293 Cells, supplied by ATCC, used in various techniques. (bioz.com)
Whole-cell voltage-clamp recordings in HEK293 cells transiently cotransfected with TRPV6 and ci-VSP were performed as described in Methods. (bioz.com)
a-d Representative traces for whole-cell patch-clamp recordings in HEK293T cells transiently cotransfected with TRPV6 and ci-VSP, performed as described in the Methods section. (bioz.com)
The atrium was mounted in an organ chamber (5 mL, Tyrode, 27 ± 0.1 ºC, 95 % O2, 5 % CO2), stretched to 10 mN, and paced at 2 Hz (0.5 ms, 400 V) and GH3 cells were submitted to a whole cell voltage clamp configuration. (scielo.br)
Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. (diavacs.com)

SSM-based electrophysiology is a suitable technique to study bacterial ion-translocating membrane- systems which are otherwise difficult to approach using conventional electrophysiological techniques, like patch clamp. (nanion.de)

For instance, free and in situ autologous tissue have been used to create a wide, unobstructed connection between the pulmonary venous confluence and the left atrium (LA). Despite this technique, 10-15% of patients represent with stenosis after initial successful correction. (medscape.com)

2. To facilitate comparison of our data with previous results and to determine the passive membrane properties under conditions as physiological as possible, recordings were made at the resting potential, in physiological saline, and without any added blockers of voltage-dependent conductances. (janelia.org)
3. Membrane-potential responses to current steps were analyzed, and four criteria were used to identify voltage responses that were the least affected by activation of voltage-dependent conductances. (janelia.org)
To evaluate the effect of the conductances underlying this sag on passive membrane properties, 2-5 mM Cs+ was added to the physiological saline. (janelia.org)

How was it gathered: We used patch clamping technique, to study the transmembrane electrical signals from individual identified astrocytes in the CA1 region of hippocampus, primary somatosensory cortex and Bergmann glia. (mendeley.com)

Although ligand binding displayed comparable K D values for muscimol among wild-type, α 1 β 2 γ 2 , and mutant receptors, patch-clamp recordings showed that α 1 β 2 (G219F)γ 2 receptors had a significantly more potent response to GABA than did α 1 β 2 γ 2 or α 1 (G223F)β 2 γ 2 . (aspetjournals.org)

In this "lab on a molecularly thin membrane", various chemical and physical operations, such as writing, erasing, functionalization and molecular transport, can be applied to user-defined regions of a membrane circuit. (nature.com)
This concept is an enabling technology for research on molecular membranes and their technological use. (nature.com)
We demonstrate how such networks are conveniently generated and locally restructured and show how various design possibilities such as diffusional barriers and hydrodynamic trapping points can be used in a "lab on a biomembrane" to directly address biomembrane functions and properties, or to perform membrane-assisted studies of molecular interactions. (nature.com)
How to use it: The raw data traces are most compatible with Clampfit 10.7.0.3 (P Clamp - Molecular devices 2016). (mendeley.com)

It currently suffers from low throughput relative to other sequencing methods mainly due to the manual labor involved in achieving a successful patch-clamp recording on a neuron. (wikipedia.org)
Investigations are currently underway to automate patch-clamp technology which will improve the throughput of patch-seq as well. (wikipedia.org)
The SURFE 2 R 96SE is the only commercially available platform for higher throughput solid supported membrane (SSM) -based electrophysiology on the market. (nanion.de)
Some researchers are even developing high-throughput techniques to track neural activity in awake, and sometimes freely moving, animals. (the-scientist.com)

Importantly, during voltage clamp recordings, histamine stimulation largely depleted the ER Ca(2+) store, explaining the activation of a CRAC-like current (due to store depletion) upon histamine in Ba(2+) medium. (unige.ch)
Cohen D, Yarom Y (1998) Patches of synchronized activity in the cerebellar cortex evoked by mossy-fiber stimulation: questioning the role of parallel fibers. (yale.edu)

Even though diverse strategies for membrane fabrication have been reported, the development of related applications and technologies has been hindered by the unavailability of both versatile and simple methods. (nature.com)

Voltage-clamp experiments validated the presence of a stretch-induced current component with a reversal potential near -11 mV. (ox.ac.uk)

Achieving this with the delicate glass tubes with incredibly thin points extending into a tip used for patch-clamp requires significant mechanical stabilization either by head fixing the animal or use of a stabilization rig attached to the animals head. (wikipedia.org)

The major theme will be devoted to describe and analyze the molecules of life, light microscopy, fine cell structure and function, cell membranes structure and function, cell division and staining procedures. (uaeu.ac.ae)

A large change in absolute ion concentrations occurs across the membrane during an action potential. (wfsahq.org)

This technique provided experimental estimates of passive membrane properties (input resistance, RN, and membrane time constant, tau m) determined in the absence of the leak conductance associated with microelectrode impalement or the washout of cytoplasmic constituents associated with conventional whole-cell recordings. (janelia.org)

Their electrical behavior was studied simultaneously in current-clamp or voltage-clamp mode using the perforated patch configuration. (ox.ac.uk)

Here we introduce a microfluidic toolbox to write 2D nanofluidic networks composed of supported phospholipid membranes and dynamically modify their connectivity, composition and local function. (nature.com)

Eventually these techniques were extended into awake, head-fixed animals and even freely moving rodents. (wikipedia.org)

Here we used direct patch-clamp of skin and eye melanosomes to identify a novel chloride-selective anion conductance mediated by OCA2 and required for melanin production. (elifesciences.org)

The most common type of albinism is caused by defects in a protein called OCA2, which is found in the membrane that surrounds melanosomes. (elifesciences.org)

This technique has been particularly useful to assess function and dysfunction of the oropharyngeal phase of swallowing. (nature.com)

Anatomy50

Organisms16

Diseases2

Chemicals and Drugs117

Analytical, Diagnostic and Therapeutic Techniques and Equipment22

Phenomena and Processes43

Disciplines and Occupations4

Technology, Industry, Agriculture4

Information Science3

Health Care1

Potentials2

Plasma membrane3

Proteins2

Cell membrane4

Currents11

Neurons10

Physiology1

Pipette3

Metabolism1

Rats1

Channels7

Biological5

Resting membrane1

Interneurons3

Lipid4

Depolarization1

Cardiac1

Bilayer1

Permeable1

Cells9

Electrophysiological1

Situ1

Conductances3

Transmembrane1

Receptors1

Molecular4

Throughput4

Stimulation2

Methods1

Experiments1

Mechanical1

Microscopy1

Concentrations1

Properties1

Simultaneously1

Connectivity1

Freely1

Direct1

Found1

Function1