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. 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. 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. However, many of the protocols and notes given in this chapter can be applied to other automated patchclamp platforms. Multiwell robotic planar patchclamp has become common in drug development and safety programmes because it enables efficient and systematic testing of compounds against. Instrumentation based on the planar patch clamp principle is made available by nanion technologies gmbh. Automated patch clamping finally achieves high throughput. For the patch clamp recordings, planar borosilicate glass patch clamp chips were used 18 in a 384microtiterplate format. Automatic patch clamp systems have been implemented in vitro using planar arrays patch onachip with microscopic orifices for seal formation fertig et al. 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. Planar patchclamp force microscopy on living cells. 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. The classical biophysics of the nerve impulse had by then been.
Automated patch clamp meets highthroughput screening. It allows you to have access to currents across a cell in two ways. Due to its dielectric properties quartz is the almost ideal material for patch clamp pipettes rae and levis, 1992. All experiments were carried out at room temperature 2224 c. The protocol described here is for the patchliner, which is provided with a tecan robot. Nanion s planar patch clamp chips offer the unique possibility to perfuse the intracellular side of the membrane. Patch clamp measurements were performed by an automated multiwell planar patch clamp system the syncropatch 384 patch engine pe nanion technologies, munich, germany fig. Thus, these techniques are not applicable for tissue slices and differentiated cells derived from ipscs or escs. An introductory guide to patch clamp electrophysiology is a concise introduction to the basic principles and practical applications of this important technique. Nanion portapatch planar patch clamping automate scientific.
The patch clamp technique wikipedia1 is widely used for the analysis of ion channels, which play an important role in many. 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 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. 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. The patchliner is an automated patch clamp robot for medium thoughput with ultimate flexibility for assay design. In addition, in planar patch clamp it is also possible to use internal agar bridges. Nanion technologies smart tools for ion channel research. The patch clamp goes planar the scientist magazine. A new technique for multiple reuse of planar patch clamp. The seal resistance was typically 12 g measurements were made with an epc 10 plus patch clamp amplifier nanion, munchen, germany. Automate scientific designs, manufactures and supplies precision biomedical instruments for neuroscience, biophysics, patch clamping, and pharmaceutical research. A single cell is then positioned on the hole by suction and a tight connection gigaseal is formed.
Copenhagen, denmarkbased sophion offers two automated planar patchclamp systems, the qpatch and the qube. Microfluidic system for planar patchclamp electrode arrays. Nanion portapatch applications automate scientific. A unique feature of nanion s automated patch clamp platforms is. The current status of automated patch clamp platforms, their strengths and drawbacks as well as. Compared with conventional patch clamp methods, the planar patch clamp method allows reliable characterization of currents from a large number of. Scientists worldwide use our automated perfusion systems, valves, amplifiers, manipulators, software, and accessories for electrophysiology, imaging, pharmacology, and drug discovery. However, many of the protocols and notes given in this chapter can be applied to other automated patch clamp platforms. Munich, germany, november 10, 2012 nanions automated patch clamp platforms are known for their high cell compatibility, even for primary cells and stem cells.
Planar patch clamp chips with a chip resistance of 23. Molecular devices, axon instrument, sophion bioscience, cytocentrics and nanion technologies have already launched their planar patch clamp systems separately. For patchliner experiments, a patchliner octo nanion technologies was used, including heka epc10 quadro amplifiers heka elektronik, lambrecht, germany. Our planar lipid bilayers are obtained from guvs prepared with the nanion vesicle prep pro setup. 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. Instead of positioning a pipette on an adherent cell, cell suspension is pipetted on a chip containing a microstructured aperture. Whole cell patch clamp recording performed on a planar glass chip. Ligand gated cng gabaa receptor glycine receptor hcn2 voltage gated hkv1. 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. Dissolve 4 mg of agar in 100 ml of 3m kcl solution by boiling and transfer to 10cm dishes. 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. 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. Does anyone use the automated patch clamp, the portapatch. 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.
Electrophysiological recordings were obtained through the use of the wholecell mode of the planar patchclamp technique portapatcho nanion, munich, germany fertig et al. Frontiers development of automated patch clamp technique. Automate scientific we build the tools so you can do the. Citeseerx document details isaac councill, lee giles, pradeep teregowda. Furthermore, the extracellular membrane remains accessible for compound application during the experiment.
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 synonyms, planar patch clamp pronunciation, planar patch clamp translation, english dictionary definition of planar patch clamp. Due to their planar chip design not only is a higher throughput achieved, but new applications have also become possible. Activation kinetics are rapid and strongly voltagedependent, whereas. Nanion s porta patch is not only the worlds smallest patch clamp setup with minimum foot print and low maintenance requirements, but. 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. Ion channels mediate electrical current flow, have crucial roles in cellular physiology, and are important drug targets. Planar patch clamping, as the name implies, is a twodimensional or planar variation on traditional patch clamping. 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.
May 24, 2006 however, patch clamping is a laborious process requiring a skilled experimenter and is, therefore, not compatible with the high throughput needed in drug development. Nanion synchropatch 384 for automated, highthroughput. Progress in automating patch clamp cellular physiology luca. First launched in 2005, the qpatch can perform from 8 to 48 simultaneous recordings from either a single cell or 10 averaged cells per site. Renaissance of ion channel research and drug discovery by. Nanion s planar patch clamp devices the porta patch is a semiautomated device with a small footprint for the analysis of ion channels in cells and lipid bilayers the patchliner is an automated patch clamp robot for medium thoughput with ultimate flexibility for assay design the syncropatch 384pe is a highthroughput patch clamp instrument which can be upgraded to the syncropatch 768pe. 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. We employ planar microstructured borosilicate glass chips nanion technologies gmbh, munich, germany, used in a commercially available patch clamp workstation porta patch from nanion, which commonly is used for automated patch clamp recording purposes alone 16,17. Nanion was nominated in 2007 for germanys most prestigious. Planar patch clamp chips in systems like the nanion porta patch are useful in that they allow more rapid throughput in drug.
Patchclamping is a goldstandard electrophysiology technique that has the temporal resolution and signal to noise ratio capable of reporting single ion channel currents, as well as electrical. Nanion s instruments use planar patch clamp chips which replace the traditional glass pipette used in. The geometry and design are covered as well as environmental manipulation techniques. Robotic multiwell planar patchclamp for native and. Planar patch clamping 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. 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. Launch of the dynamic clamp unit for the patchliner the. Microchip technology for automated and parallel patch. Armin kargol statement about the portapatch nanions portapatch is an excellent instrument for research labs working with undergraduate students.
Levis and rae, 1993 and is thus a very suitable substrate for planar patch clamp chips. Nanion s planar patch clamp chips for the porta patch not only allow for recordings of cell membranes but also of artificial bilayers. 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. 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 cell membrane 30. Automated electrophysiological and pharmacological evaluation. Nanion s instruments use planar patch clamp chips which replace the traditional glass pipette used in the technique of patch clamping. 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. 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.
Gigaseal patch clamping 384 or 768 recording channels up to 20,00040,000 data points per day unattended operation 46 hours 0,16 0,65 per data point ultra flexible. The cells are sealed to the substrate and either the whole cell configuration is reached by application of suction pulses, or poreforming compounds are applied for perforated patch recordings allowing electrical access and control of the cellular membrane. A new technique for multiple reuse of planar patch clamp chips. As in conventional patch clamp, agar bridges are used for the reference electrode. 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. 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. Nanion was nominated in 2007 for germanys most prestigious innovation award the deutscher zukunftspreis german future prize, federal presidents award for technology and innovation. Manual patchclamping methods depend to a large extend on the skills of the operator. Nanion and axiogenesis present parallel patch clamp. Nanion vesicle prep pro bilayer apparatus automate. Nanion introduces the orbit 16 a pioneering device for.
Traditional patch clamp has a very low throughput which has been proven to be a. 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. This aspect is exploited with the internal perfusion system for the portapatch. Development of planar patch clamp technology and its. Commercial systems available for multiwell planar patch clamp include patchxpress 7000a. 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. A comparison of the performance and application differences between manual and automated patch clamp techniques. Planar patch clamp definition of planar patch clamp by the. Dec 17, 2008 nanion s instruments use planar patch clamp chips which replace the traditional glass pipette used in the technique of patch clamping. 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.
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. Ion channel screening automated patch clamp on the rise. 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. A benchmark study with seal chip planar patchclamp. Planar patch clamp approach to characterize ionic currents. Since it was founded in 2002, nanion has grown to one of the most respected automated patch clamp providers worldwide. Highthroughput electrophysiology with planar patch clamping.
With its vast experimental freedom and gigaseal data quality, the patchliner is one of the most versatile patch clamp instruments on the market. 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. Porta patch planar patch clamping the porta patch is a onecellatatime system, which is surprisingly small for a complete patch clamp rig. Oct 23, 2018 a whole cell patch clamp measures currents through the entire cell membrane.
Planar patch clamp is a novel method developed for high throughput electrophysiology. Cell suspensions for apc devices utilizing planar patch clamp chips should be homogeneous milligan et al. The planar geometry of the chip is predestined for the combination with an afm. The advent of automated patch clamp platforms has revolutionized ion channel screening, enabling investigations from a more functional perspective at a much higher throughput.
It is carried out similarly to cell attached patch clamping, but more suction is applied in order to rupture the membrane. A patch clamp recording of current reveals transitions between two conductance states of a single ion channel. Robotic multiwell planar patchclamp for native and primary. Ion channels are integral membrane proteins that regulate the flow of ions across the plasma membrane. Planar patch clamps planar patch clamping, as the name implies, is a twodimensional or planar variation on traditional patch clamping. Experiments are performed on a single cell at a time, whereby whole dose response curves can be obtained from a single cell. Planar patch clamp definition of planar patch clamp by. It has a much smaller foot print compared to a conventional rig. We employ planar microstructured borosilicate glass chips nanion technologies gmbh, munich, germany, used in a commercially available patch clamp workstation porta patch from nanion, which commonly is used for automated patch clamp recording purposes alone.
Ion channels are vital to a wide variety of biological processes and are prominent components of the nervous system and. The channel activity is induced upon membrane depolarization to voltages more positive than 40 mv. Nanion develops and markets instrumentation patch clamp devices, related consumables for instance, patch plates, and services for automated patch clamping. For planar patch clamp, a wide range of different cell suspensions can be used bruggemann et al. The device consists of a planar chip made from fused quartz. Patchliner versatile and sophisticated the patchliner is a fully automated planar patch clamp instrument recording from up to 8 cells simultaneously. Using optimized conditions, we recorded both wt and corrected f508delcftr chloride currents with automated whole cell patch clamp and compared the data to results.
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