Microchip technology for automated and parallel patch. A single cell is then positioned on the hole by suction and a tight connection gigaseal is formed. From understanding cellular function to novel drug. The tip of the pipette figure figure1a 1 a is replaced by a micronsized aperture through a selfsupported thin film figure figure1b 1 b separating an upper cell suspension chamber from an underlying pipette. However, mass patch clamp techniques, such as planar patch clamp 7,8 or population patch clamp 9,10 using labsonachip and other microfluidic devices 11 are usually applied for the. This aspect is exploited with the internal perfusion system for the portapatch. Patch clamp recordings in the whole cell configuration, and single channel recordings in the cell attached configuration, can be performed with planar patch clamp chips. For instance, scanning probe techniques such as force microscopy or near field optical microscopy can easily be performed on the planar patch clamp chips presented here. A mpc involves the use of a glass pipette that is manoeuvred to an adherent cell approach. The planar patch clamp technique overcomes some of the major limitations of the conventional patch clamp, while still offering the same advantages, such as temporal resolution, voltage control and direct measurement of a compounds mode of action. Planar patch clamp definition of planar patch clamp by the. Tedious manual patch clamp rigs have given way to automated planar patch clamp systems with sufficient throughput to play a role in the drugdevelopment process.
Ion channels are integral membrane proteins that regulate the flow of ions across the plasma membrane. A new technique for multiple reuse of planar patch clamp chips. The portapatch from nanion is a platform close to manual patch clamp. A unique feature of nanion s automated patch clamp platforms is the possibility to exchange intracellular components. Development of planar patch clamp technology and its application in. For many years now manual patchclamping has been regarded as one of the best. Here we report on a planar, microstructured quartz chip for whole cell patch clamp measurements without micromanipulation or visual control. Experiments are performed on a single cell at a time, whereby whole dose response curves can be obtained from a single cell. Fortunately, automation has entered the area of experimental electrophysiology over the past few years. He discovered action potential or negative variation, as it was called at the time, one of the main types of electric processes in excitable tissues.
Novel screening techniques for ion channel targeting drugs. For patchliner experiments, a patchliner octo nanion. Automated patch clamping finally achieves high throughput. Instrumentation based on the planar patch clamp principle is made available by nanion technologies gmbh.
In 2009 nanion was awarded the german founders award grunderpreis, and the innovation prize step award. Patch clamping is still a laborious process, requiring a skilled experimenter to micromanipulate a glass pipette under a microscope to record from one cell at a time. Using automated patch clamp electrophysiology platforms in. We request funding to purchase a nanion synchropatch planar patch clamp electrophysiology system to alleviate the bottlenecks that slow the pace of ion channel small molecule discovery and potentially hasten the development of new therapeutics. Portapatch planar patch clamping the portapatch is a onecellatatime system, which is surprisingly small for a complete patch clamp rig. This type of clamp forms a looseseal interaction between the cell membrane and the rim of the glass microelectrode. Nanion synchropatch 384 for automated, highthroughput.
Dec 28, 2012 the gigaohm seal can be achieved and whole cell access or other patch clamp configurations may be achieved in the same fashion as the conventional manual patch clamp technique. We employ planar microstructured borosilicate glass chips nanion technologies gmbh, munich, germany, used in a commercially available patchclamp workstation portapatch from nanion, which commonly is used for automated patchclamp recording purposes alone 16,17. To date, a number of planar patch clamp systems have been developed, ranging from small devices recording from a single cell at a time, e. Munich, germany, november 10, 2012 nanions automated patch clamp platforms are known for their high cell compatibility, even for primary cells and stem cells. The patch clamp technique is the stateoftheart technology for the study of ion channels, but patch clamping is a laborious process requiring a skilled and highly trained scientist. Nanion technologies gmbh manufactures medical appratus. Frontiers development of automated patch clamp technique. A comparison of the performance and application differences. Unlike other planar patch clamp devices we considered, the patchliner gave us full access to the electrophysiology modes we require for a broad range of experimental protocols including voltage clamp, current clamp, standard whole cell, cell attached, and perforated patch configurations combined with a facile ability to exchange the internal solution. Nanions planar patch clamp devices the portapatch is a semiautomated device with a small footprint for the analysis of ion channels in cells and lipid bilayers. Most platforms utilize borosilicate glass configured as a pipette, however more recently planar patch clamp chips have been developed that require less technical expertise. The company offers wide range of patch clamp devices, techniques, and electro physiological products. Nanion portapatch planar patch clamping automate scientific.
Traditional patch clamp has a very low throughput which has been proven to be a. Whole cell patch clamp recording performed on a planar glass. Dec 17, 2008 nanion s instruments use planar patch clamp chips which replace the traditional glass pipette used in the technique of patch clamping. This technique also has the ability to perform rapid.
The planar method of patchclamp has special features that need to be handled by the investigator. Nanion technologies and data analysis datacontrol 384, nanion technologies. Planar patch clamp chips in systems like the nanion portapatch are useful in that they allow more rapid throughput in drug screening studies. The patchliner is an automated patch clamp robot for medium thoughput with ultimate flexibility for assay design. A benchmark study with seal chip planar patchclamp. For the patch clamp recordings, planar borosilicate glass patch clamp chips were used18 in a 384microtiterplate format. Here we describe features of nanions npc16 patchliner and processes and protocols suited for this particularly flexible and successful highthroughput automated platform, which is based on planar patchclamp technology. Due to their planar chip design not only is a higher throughput achieved, but new applications have also become possible. Nov 07, 20 tedious manual patchclamp rigs have given way to automated planar patchclamp systems with sufficient throughput to play a role in the drugdevelopment process. Planar patch clamp definition of planar patch clamp by. Usually small glass pipettes are used for the electrical analysis or manipulation of the cell. Planar patch clamping, as the name implies, is a twodimensional or planar variation on traditional patch clamping. Jun 08, 2012 whole cell patch clamp recordings from morphologically digitimer ds2a duration. Nov 10, 2015 to date, a number of planar patch clamp systems have been developed, ranging from small devices recording from a single cell at a time, e.
Robotic multiwell planar patchclamp for native and primary mammalian cells carol j milligan 1, jing li 1, piruthivi sukumar 1, yasser majeed 1, mark l dallas 2, anne english 3, paul emery 3. For example, some membrane processes may be adversely affected by the protocol required to produce the cell suspension, or by seal enhancer solutions. The setup not only allows for immobilizing even a nonadherent cell for measurements of its mechanical properties, but also for simultaneously measuring the electrophysiological properties of a single cell. Oct 23, 2018 the patch clamp is a laboratory technique for studying currents in living cells. The technique of patch clamping can be seen in retrospect as a combination of two separate lines of development that both originated in the 1960s and 1970s. The patch clamp technique is a laboratory technique in electrophysiology used to study ionic currents in individual isolated living cells, tissue sections, or patches of cell membrane. Highthroughput electrophysiology with planar patch clamping. Nanion synchropatch 384 for automated, highthroughput electrophysiology. These planar patch clamp systems have properties of a high throughput, the automatic formation of a high seal resistance, the involvement of voltage and current clamps, and the capability of wholecell and single channel recordings, drug perfusion and data analysis. The patch clamp goes planar the scientist magazine. Does anyone use the automated patch clamp, the portapatch. Situated in the heart of munich, nanion is vibrant team of scientists and engineers with the goal of providing top quality instrumentation and customer service for a broad range of analytical applications. Many platforms currently use the planarpatch technology to achieve.
It helped me a lot when i performed potch a patch system. Npc 16 patchliner and syncropatch 96 nanion technologies gmbh, munich. Planar patchclamp force microscopy on living cells. Development of planar patch clamp technology and its. A single ion channel conducts around 10 million ions per second. The microelectrode pipette, whose aperture provides the suction that couples the instrument to the cell, is replaced by a planar electrode substrate into which a hole has been bored. Automated patch clamp increases throughput and ease of use compared to the conventional patch clamp technique making it accessible to a wider audience. With conventional patch clamp this is difficult because of the tiny dimensions of the pipette tip, the long tapered shaft and, thus. Renaissance of ion channel research and drug discovery by. These planar patch clamp systems have properties of a high throughput, the automatic formation of a high seal resistance, the involvement of voltage and current clamps, and the. Contact is then made with the cell, and through application of negative pressure, an electrical seal forms, in the order of g. We utilized the novel wholecell mode of the planar patch clamp technique porta patch nanion, munich, germany394041 42 to detect wholecell currents in bon1 cells. Finally, transferring the patch clamp technique onto a planar device enables a variety of new kinds of experiments on ion channels.
In this study, we evaluated the use of planar parallel apc technique for. Frontiers development of automated patch clamp technique to. Several companies have released or are planning to release commercial implementations of a new highthroughput variation on the technique, called planar patch clamping. The classical biophysics of the nerve impulse had by then been established in the squid giant axon using a combination of 1 voltage clamping with axial wire electrodes and 2 internal perfusion or dialysis. In addition, in planar patch clamp it is also possible to use internal agar bridges. Both processes need to be understood to achieve consistent delivery of the desired compound into the cell. The paradigm shift from cellattached mode in oocytes to wholecell mammalian planar patch clamp was first published by niels fertig and colleagues at nanion. Planar patch clamp is a novel method developed for high throughput electrophysiology.
The device consists of a planar chip made from fused quartz. The patch clamp technique is widely used for recording the activity of ion channels in single cells and lipid bilayers. Following the successful market introduction of two automated patch clamp devices, the porta patch 2004 and the patchliner 2006, nanion now introduces a massively parallel patch clamp platform, the. These new technologies are designed to overcome many of the hurdles associated with traditional wholecell voltage clamp by dramatically reducing time and expertise required to perform. These planar patch clamp systems have properties of a high throughput, the automatic formation of a high seal resistance, the involvement of voltage and current clamps, and the capability of wholecell and single channel recordings, drug. The loose patch clamp is different from the patch clamps discussed above. The planar patch clamp technique allows only two configurations. Patchliner versatile and sophisticated the patchliner is a fully automated planar patch clamp instrument recording from up to 8 cells simultaneously. One of the advantages of planar patch clamp is the possibility of perfusing the intracellular side of the membrane during a patch clamp experiment in the wholecell configuration. However, many of the protocols and notes given in this chapter can be applied to other automated patchclamp platforms.
This technique also has the ability to perform rapid solution changes from the intracellular side. Here, pore forming molecules are used to perforate the membrane patch in order to obtain electrical control over the membrane. Instead of a micromanipulator and manually positioned electrodes, planar patch clamps query cells by sucking them onto tiny holes in the bottom of custom microtiter plates. Using optimized conditions, we recorded both wt and corrected f508delcftr chloride currents with automated whole cell patch clamp and compared the data to results.
Owing to the ultralowthroughput, conventional patch clamping is frequently applied only in the later stages of drug discovery and development. Microfluidic system for planar patchclamp electrode arrays xiaohui li yale university 2006 the patch clamp has been widely accepted as a standard technique for fundamental studies of ion channel proteins, and discovery of drugs that affect these proteins. It is usually carried out by applying a voltage across the cell membrane and measuring the resulting current. Data output and compound information is compatible with most database formats. In a second approach, a tiny pipette tip is vertically positioned upwards in the center of a polyimide sheet that is set at the bottom of a well. The technique of patch clamping can be seen in retrospect as a combination of two separate lines of development that both origi nated in the 1960s and 1970s. Nowadays, the planar patch clamp systems are being developed toward miniaturization and portability, e. It has a much smaller foot print than the conventional rig. Citeseerx document details isaac councill, lee giles, pradeep teregowda. Planar patch clamp approach to characterize ionic currents. Whole cell patch clamp recording performed on a planar. In these automatic devices like the planar population array patch clamp, papc, the micropipettes are replaced by pores in planar supports. Nov 30, 2011 nanions high quality instruments employ planar patch clamp chips which replace the traditional glass pipette used in the conventional patch clamp technique. Microfluidic system for planar patch clamp electrode arrays xiaohui li yale university 2006 the patch clamp has been widely accepted as a standard technique for fundamental studies of ion channel proteins, and discovery of drugs that affect these proteins.
Patch clamp recordings in the whole cell configuration, and single channel recordings in the cell attached configuration, can be performed with planar patch. A patch clamp recording of current reveals transitions between two conductance states of a single ion channel. Planar patch clamp article about planar patch clamp by the. Molecular devices, axon instrument, sophion bioscience, cytocentrics and nanion technologies have already launched their planar patch clamp systems separately. Nanions syncropatch 96 convinces big pharma automated patch. The technique is especially useful in the study of excitable cells such as neurons, cardiomyocytes, muscle fibers, and pancreatic beta cells, and can also be applied to the study of bacterial ion channels in. A planar patchclamp chip microstructured from borosilicate glass was used as a support for mechanical probing of living cells. The combination of a patch clamp setup with an ultrasonic system overcomes this obstacle and allows the study of sonoporation as well as the resealing of the pores in the cell membrane in realtime. Nanion s planar patch clamp chips offer the unique possibility to perfuse the intracellular side of the membrane.
As in conventional patch clamp, agar bridges are used for the reference electrode. Planar patch clamp synonyms, planar patch clamp pronunciation, planar patch clamp translation, english dictionary definition of planar patch clamp. Ion channel screening automated patch clamp on the rise. Dissolve 4 mg of agar in 100 ml of 3m kcl solution by boiling and transfer to 10cm dishes. Microfluidic system for planar patchclamp electrode arrays. With its vast experimental freedom and gigaseal data quality, the patchliner is one of the most versatile patch clamp instruments on the market. In this study we evaluated the use of planar parallel automated patch clamp technique for pharmacological search of cftr trafficking correctors and cftr function modulators. It is thus of special interest in the research of excitable cells such as neurons, cardiomyocytes and muscle fibers. The wholecell mode of the planar patchclamp technique portapatcho nanion, munich. Since it was founded in 2002, nanion has grown to one of the most respected automated patch clamp providers worldwide.
Few works used automated patch clamp apc for cftr recording, and none have. The patch clamp technique allows the investigation of a small set or even single ion channels. Automated patch clamp meets highthroughput screening. There is a book patch clamp analysis advanced techniques, chapter 14 planar patch clamping, which i think would help you. Robotic multiwell planar patchclamp for native and. The portapatch from nanion is a platform close to manual patchclamp. With conventional patch clamp this is difficult because of the tiny dimensions of the pipette tip, the long tapered shaft and, thus, the diffusion limitation of the liquid exchange. Duboisreymond, using a more advanced technique, confirmed that an injury to a muscle or nerve invariably results in a difference in potentials that decreases upon excitation. Patchlinernpc16 workstation nanion technologies gmbh. A significant advantage of this technique is that the pipette can be used repeatedly and the cell membrane remains intact. Nanion portapatch applications automate scientific. A new technique for multiple reuse of planar patch clamp. Highthroughput electrophysiology with planar patch. Planar patch clamp chips have been developed and refined over the last decade to specifically address the above limitation sigworth and klemic, 2005.
Increasing the temperature to over 40 c induced reversible rises in nonselective cation currents. Request pdf a new technique for multiple reuse of planar patch clamp chips the patch clamp technique is widely used for recording the activity of ion channels in single cells and lipid bilayers. Whole cell patch clamp recording performed on a planar glass chip. For automated high throughput screening the concept of planar patchclamp was developed recently 10. Instead of positioning a pipette on an adherent cell, cell suspension is pipetted on a chip containing a microstructured aperture. Nanion s porta patch, claims to be the worlds smallest patch clamp workstation, and uses nanion s planar patch clamp chips providing data on any ion channel target. The automation of the patch clamp method was made possible with the development of the chipbased planar patch clamp technology. Levis and rae, 1993 and is thus a very suitable substrate for planar patch clamp chips. Nanion has been the first on the market with the launch of the semiautomated planar patch clamp device, the porta patch, in 2003. Nanions planar patch clamp chips offer the unique possibility to perfuse the. Ion channels mediate electrical current flow, have crucial roles in cellular physiology, and are important drug targets. The automation of the manual patchclamp technique involved an evolution of innovative steps.
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