For the patch clamp recordings, planar borosilicate glass patch clamp chips were used 18 in a 384microtiterplate format. Frontiers development of automated patch clamp technique to. Electrophysiological analysis of mammalian cells expressing. Nanion was nominated in 2007 for germanys most prestigious innovation award the deutscher zukunftspreis german future prize, federal presidents award for. The patchliner is a fully automated planar patch clamp. Patch clamp electrophysiology has transformed research in the life sciences. Development of planar patch clamp technology and its. Building on the success of this instrument, the patchliner a fully automated patch clamp platform was launched in 2006. The chloride cl channel cystic fibrosis transmembrane conductance regulator cftr is defective in cystic fibrosis cf, and mutation of its encoding gene leads to various defects such as retention of the misfolded protein in the endoplasmic reticulum, reduced stability at the plasma membrane, abnormal channel gating with low open probability, and thermal instability, which leads to.
Ion channels mediate electrical current flow, have crucial roles in cellular physiology, and are important drug targets. Patch clamp recordings in the whole cell configuration, and single channel recordings in the cell attached configuration, can be performed with planar patch. Planar patch clamp definition of planar patch clamp by. The highthroughput system used here is a planar patch clamp screening. Not only a powerful research tool but also ideal for educational purposes and quick tests of cells and ion channels. Planar patch clamp chips with a chip resistance of 23. Npc16 patchliner and syncropatch 96 nanion technologies gmbh. Whole cell patch clamp recording performed on a planar. Nanion s planar patch clamp chips offer the unique possibility to perfuse the intracellular side of the membrane.
Instead of positioning a pipette on an adherent cell, cell suspension is pipetted on a chip containing a microstructured aperture. Nanions syncropatch 96 convinces big pharma automated patch. Planar patch clamp chips in systems like the nanion porta patch are useful in that they allow more rapid throughput in drug. 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. Bleach solution nanion sodium hypochlorite solution naclo, 12% cl cacl 22h 2o camsa. Nanion technologies and data analysis datacontrol 384, nanion technologies. 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.
One of these breakthroughs is the development of automated planar patchclamp. Progress in automating patch clamp cellular physiology luca a. Frontiers stateoftheart automated patch clamp devices. A new technique for multiple reuse of planar patch clamp. Nanion portapatch planar patch clamping automate scientific. For many years now manual patchclamping has been regarded as one of the. Nanion and axiogenesis present parallel patch clamp.
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. 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. Whole cell patch clamp recording performed on a planar glass chip. Conventional patchclamp involves the maneuvering, under a microscope, of the fine tip 12. Nanion vesicle prep pro bilayer apparatus automate. Nanion synchropatch 384 for automated, highthroughput. Munich, germany, november 10, 2012 nanions automated patch clamp platforms are known for their high cell compatibility, even for primary cells and stem cells. Data output and compound information is compatible with most database formats.
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. Planar patch clamp synonyms, planar patch clamp pronunciation, planar patch clamp translation, english dictionary definition of planar patch clamp. A unique feature of nanion s automated patch clamp platforms is the possibility to exchange intracellular components. Automated patch clamp meets highthroughput screening.
Using optimized conditions, we recorded both wt and corrected f508delcftr chloride currents with automated whole cell patch clamp and compared the data to results. Planar patch clamping, as the name implies, is a twodimensional or planar variation on traditional patch clamping. Automated electrophysiological and pharmacological. Planar patch clamp is a novel method developed for high throughput electrophysiology. The planar lipid bilayers can be prepared on the chip either by the painting method or by guv giant unilammellar vesicleliposome fusion on the aperture. A new technique for multiple reuse of planar patch clamp chips. The wholecell mode of the planar patchclamp technique portapatcho nanion, munich. All experiments were carried out at room temperature 2224 c. Portapatch planar patch clamping the portapatch is a onecellatatime system, which is surprisingly small for a complete patch clamp rig. Nanion technologies was founded in 2002 as a spinoff from the university of munich, center for nanoscience cens, where niels fertig undertook his phd developing planar chip substrates for patch clamp experiments on cells and lipid bilayers. Automated patch clamping finally achieves high throughput. Robotic multiwell planar patchclamp for native and primary. A single cell is then positioned on the hole by suction and a tight connection gigaseal is formed. Using automated patch clamp electrophysiology platforms in.
Planar patch clamp devices, such as the three described in this unit, are overtaking glass pipets in popularity because they increase throughput, are easier to use, provide for the acquisition of high. Nanions syncropatch 384768pe is a high throughput planar patchclamp platform that achieves high and robust success rates. Compared with conventional patch clamp methods, the planar patch clamp method allows reliable characterization of currents from a large number of isolated lysosomes up to eight lysosomal recordings per preparation. Nanions planar patch clamp chips offer the unique possibility to perfuse the intracellular side of the membrane. Ion channels have gained increased interest as therapeutic targets over recent years, since a growing number.
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. Planar patch clamp chips in systems like the nanion portapatch are useful in that they allow more rapid throughput in drug screening studies. Citeseerx document details isaac councill, lee giles, pradeep teregowda. This protocol assumes basic knowledge of patch clamp methods. Aug 07, 2015 384 or 768 cells recorded in parallel 20,000 data points per day 8595 % success rates routinely obtained reliable data from diverse ion channels targets seamless integration into. Frontiers development of automated patch clamp technique. The syncropatch 384 pe was a patchclamp module that was integrated into a liquid handling robot owning a 384 pipetting head, biomek fx beckman coulter, brea, ca, usa. Microfluidic system for planar patchclamp electrode arrays. This aspect is exploited with the internal perfusion. May 22, 2005 nanion s product is based on fertigs research, which bezanilla and other scientists praise highly. The portapatch is a miniaturized patch clamp system supporting gigaseal recordings from one cell at a time.
Nanions instruments use planar patch clamp chips which replace the traditional glass pipette used in the technique of patch clamping. 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. Nowadays, the planar patch clamp systems are being developed toward miniaturization and portability, e. The most popular whole cell variant of the technique detects the ensemble current over the entire cell membrane. For planar patch clamp, a wide range of different cell suspensions can be used bruggemann et al. Dec 16, 2015 patch clamp measurements were performed by an automated multiwell planar patch clamp system the syncropatch 384 patch engine pe nanion technologies, munich, germany fig.
Ion channels are integral membrane proteins that regulate the fl ow of ions across the plasma membrane and the membranes of intracellular organelles of both excitable and nonexcitable cells. Planar patch clamp recording allows for high throughput and parallel invitro recording by using a planar device with holes made of silicon or glass to create the seal with the cells. In conventional patch clamping the cell is accessed by means of a glass pipette that is manually brought into contact with its membrane 38, while in planar patch clamp the traditional glass pipette is replaced by a hole at the bottom of a well through which an applied negative pressure patches the. The geometry and design are covered as well as environmental manipulation techniques. Nanion combines bio and nanotechnology in a company serving the life sciences industry by offering products and services that increase the speed and efficiency of the drug discovery process. Molecular devices, axon instrument, sophion bioscience, cytocentrics and nanion technologies have already launched their planar patch clamp systems separately. Whole cell patch clamp recording performed on a planar glass. Nanion bases its business on a proprietary chip technology and. Planar patchclamp recording allows for high throughput and parallel invitro recording by using a planar device with holes made of silicon or glass to create the seal with the cells. Nanions high quality instruments employ planar patch clamp chips which replace the traditional glass pipette used in the conventional patch clamp technique. Configuration of suction and voltage parameters required for seal formation and membrane rupture in planar lysosomal patch clamp. We here report the development of an automatic device for whole cell patch clamping that can easily be scaled up into a parallel array. Planar patch clamp approach to characterize ionic currents.
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. Cell suspensions for apc devices utilizing planar patch clamp chips should be homogeneous milligan et al. The patchliner is an automated patch clamp robot for medium thoughput with ultimate flexibility for assay design. 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. A unique feature of nanions automated patch clamp platforms is the possibility to exchange intracellular components. During a patch clamp recording, a hollow glass tube known as a micropipette or patch pipette filled with an electrolyte solution and a recording electrode connected to an amplifier is brought into contact with the membrane of an isolated cell. It allows you to have access to currents across a cell in two ways. 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. Robotic multiwell planar patchclamp for native and. For patchliner experiments, a patchliner octo nanion. Using automated patch clamp electrophysiology platforms in painrelated ion channel research. Nanion is a leading provider of automated patch clamp apc electrophysiology systems with throughput capabilities ranging from a single cell up to fully automated high throughput screening hts instruments recording from 384 cells simultaneously. Planar patch clamp definition of planar patch clamp by the.
Clearly, the planar patch clamp has been a facilitating technology, which significantly improved ion channel screening by increasing the throughput and easeofuse of electrophysiology in drug discovery. The patchliner is a fully automated planar patch clamp instrument recording from up to 8 cells simultaneously. 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. With this method, the glass pipette of conventional patchclamp recording is replaced by a thin sheet of flat glass with a small aperture in the middle the planar chip. A planar patch clamp device with an accessible aperture is ideally suited for these kinds of combined experiments by which new insights on ion channel behavior can be gained. Planar patch clamp approach to characterize ionic currents from. 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. 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. Nanion ups throughput and cuts costs with a new industrial. As in conventional patch clamp, agar bridges are used for the reference electrode. Microchip technology for automated and parallel 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. The patch clamp goes planar the scientist magazine.
For the patch clamp recordings, planar borosilicate glass patch clamp chips were used18 in a 384microtiterplate format. Nanion vesicle prep pro bilayer apparatus automate scientific. The syncropatch 384 pe was a patch clamp module that was integrated into a liquid handling robot owning a 384 pipetting head, biomek fx beckman coulter, brea, ca, usa. Nano patchclamp npc development of a highthroughput system based on planar patchclamp patch clamping is a marvelous technique. 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. Patchclamp measurements were performed by an automated multiwell planar patchclamp system the syncropatch 384 patch engine pe nanion technologies, munich, germany fig. Contact is then made with the cell, and through application of negative pressure, an electrical seal forms, in the order of g. Our first patch clamp device, the portapatch the smallest patch clamp rig in the world, was launched in 2003. Development of planar patch clamp technology and its application in.
Automated patchclamp system patchliner nanion technologies. Planar patch clamp synonyms, planar patch clamp pronunciation, planar patch clamp translation, english dictionary definition of planar. We have found that robotic multiwell planar patchclamp technology can generate high quality publishable data1012 and illustrative data shown in this article from native cells and cells in primary culture, as well as confer advantages and be enabling compared with conventional patchclamp. Experiments are performed on a single cell at a time, whereby whole dose response curves can be obtained from a single cell. 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. Ion channels are vital to a wide variety of biological processes and are prominent components of the nervous system and. High gigaseal rate on hek, cho, ltk, rbl, ips and rbc. In 2009, nanion lauched its high throughput, highquality automated patch clamp system, the syncropatch 96. In addition, in planar patch clamp it is also possible to use internal agar bridges.
For patchliner experiments, a patchliner octo nanion technologies was used, including heka epc10 quadro amplifiers heka elektronik, lambrecht. Patch clamping is still a laborious process, requiring a skilled. Dissolve 4 mg of agar in 100 ml of 3m kcl solution by boiling and transfer to 10cm dishes. The state of the art technology for the study of ion channels is the patch clamp technique. Renaissance of ion channel research and drug discovery by.
Levis and rae, 1993 and is thus a very suitable substrate for planar patch clamp chips. With its vast experimental freedom and gigaseal data quality, the patchliner is one of the most versatile patch clamp instruments on the market. The system uses nanions planar patch clamp chips and generates high quality data. Following the successful market introduction of two automated patch clamp devices, the portapatch 2004 and the patchliner 2006, nanion now introduces a massively parallel patch clamp platform, the. Nanions planar patch clamp chips for the portapatch not only allow for recordings of cell membranes but also of artificial bilayers. Traditional patch clamp has a very low throughput which has been proven to be a. Instrumentation based on the planar patch clamp principle is made available by nanion technologies gmbh. More than 10 years experience in assay design and support. However, many of the protocols and notes given in this chapter can be applied to other automated patchclamp platforms. Nanions portapatch is not only the worlds smallest patchclamp setup with minimum foot print and low maintenance requirements, but also the cheapest way to access apc technology. Ampio pharmaceuticals announces agreement to acquire key delivery technology. This aspect is exploited with the internal perfusion system for the portapatch. Due to their planar chip design not only is a higher throughput achieved, but new applications have also become possible.
Planar patch clamp chips in systems like the nanion portapatch are useful in that they allow more rapid throughput in drug. Nanions product is based on fertigs research, which bezanilla and other scientists praise highly. 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. Due to its dielectric properties quartz is the almost ideal material for patch clamp pipettes rae and levis, 1992. Highthroughput automated patch clamp complemented by.
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. A mpc involves the use of a glass pipette that is manoeuvred to an adherent cell approach. Nanions instruments use planar patch clamp chips, which replace the traditional glass pipette used in the technique of patch clamp electrophysiology. It offers fast and easy access to high quality patch clamp data with only minimal training.