Ion channel screening automated patch clamp on the rise. Thus, these techniques are not applicable for tissue slices and differentiated cells derived from ipscs or escs. Apr 17, 2020 nanion technologies is a spinoff from the center of nanoscience cens of the university of munich lmu. Frontiers stateoftheart automated patch clamp devices. Cell suspensions for apc devices utilizing planar patch clamp chips. One of the advantages of planar patch clamp is the possibility of perfusing the intracellular side of the membrane. Several companies have released or are planning to release commercial implementations of a new highthroughput variation on the technique, called planar patch clamping. How patch clamp technology can benefit ion channel research. Oocyte is confined in a cone shaped well to a fixed position.
Instrumentation based on the planar patch clamp principle is made available by nanion technologies gmbh. I worked 6 years ago with the portapatch device and we had no problems to. Up to eight different solutions can be perfused inside the cell, allowing dose response analysis of compounds acting on the cytosolic portion of the ion channel. The characterization of drug effects on human currents using automated patch clamp apc platforms employing single or multihole planar patch techniques 1,2,3,4,5 has revolutionized the. Robotic multiwell planar patchclamp for native and primary. Automated planararray electrophysiological platforms. The portapatch is a miniaturized patch clamp system supporting gigaseal recordings from one cell at a time.
Here we describe features of nanion s npc16 patchliner and processes and protocols suited for this particularly flexible and successful highthroughput automated platform, which is based on planar patch clamp technology. Unlike other planar patch clamp devices we considered, the patchliner gave us. The patch clamp technique allows the investigation of a small set or even single ion channels. 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. Feb 23, 2015 patch clamp recordingpatch clamp recording the patch clamp technique is a laboratory technique in electrophysiology that allows the study of single or multiple ion channels in cells. The first device was fabricated in silicon fertig et al. 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. Nanion s high quality instruments employ planar patch clamp chips which replace the traditional glass pipette used in the conventional patch clamp technique. 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. 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.
We miniaturized this patch clamp system even further by integrating the amplifier into the system. 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. Analysis advanced techniques, chapter 14 planar patch clamping, which i. Nanion portapatch applications automate scientific. A successful robotic approach of the patch clamp technique is based on planar patch clamp chips where a glass pipette, as used in conventional patch clamping, is replaced by a thin planar glass. Patchclamp setup conventional patch clamping, now also referred to as manual patchclamp to distinguish it from the recently developed automated patchclamp described below see section vi, is accomplished by sealing the small tip of a pipette to the surface of the cell membrane in such a way that is possible to isolate a tiny. Reversible control of current across lipid membranes by local. Ion channels are essential in a wide range of cellular functions and their malfunction underlies many disease states making them important targets in drug discovery. Planar patch clamp combined with atomic force microscope afm. It is comparatively easy to obtain a good cellattached seal in patch clamp using oocytes, but most electrophysiology is performed in mammalian cells in wholecell mode. How patch clamp technology can benefit ion channel.
The present study aimed to evaluate the nanion patchliner. Molecular devices, axon instrument, sophion bioscience, cytocentrics and nanion technologies have already launched their planar patch clamp systems separately. Nanions instruments use planar patch clamp chips, which replace the traditional glass pipette used in the technique of patch clamp electrophysiology. The advent of automated patch clamp platforms has revolutionized ion channel screening, enabling investigations from a more functional perspective at a much higher throughput. Aug 19, 2019 the porta patch mini is built on the success of the porta patch. A recent explosion in the number and different approaches taken to planar array patch clamp of mammalian cells has occurred in the past few years, resulting in the commercialization of four planar. Due to their planar chip design not only is a higher throughput achieved, but new applications have also become possible. The ionworks ht also is highthroughput, allowing planar patch clamp testing in 384 wells at once. It is thus of special interest in the research of excitable cells such as neurons, cardiomyocytes and muscle fibers. Continuing the research of the bestselling first edition, patch clamp analysis. However, many of the protocols and notes given in this chapter can be applied to other automated patch clamp platforms.
Select any or all components you need for your custom electrophysiology rig. Experiments were performed on a commercial setup for planar patch clamp nanion porta patch. The paradigm shift from cellattached mode in oocytes to wholecell mammalian planar patch clamp was first published by niels fertig and colleagues at nanion. Since the volume of perfusion chambers oocyte well is only 4 microliter, the solution change is almost instantaneous. 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. May 22, 2005 the ionworks ht also is highthroughput, allowing planar patch clamp testing in 384 wells at once. It is a research grade instrument allowing for whole cell as well as single channel recordings with great experimental versatility. All automated patch clamping ezgseal 100b pressure controller the ezgseal 100b pressure controller is designed mainly for automated patch clamping, but it can also be used for many other applications. Patch clamp electrophysiology has transformed research in the life sciences over the last few decades. Nanions syncropatch 96 convinces big pharma automated patch. 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. I have used patchliner ordered from nanion, but the success rate is not as high. 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. This project focuses on the simultaneous measurements of ionchannels using ligand tethered afm tips combined with a planar patch clamp device.
Ionflux automated hts patch clamp system mercury and mercury ht blm planar lipid bilayer workstation. Atomic force microscopy and patch clamp are two incompatible techniques as long as patch clamp will be routinely done using glass pipettes. A new technique for multiple reuse of planar patch clamp. 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. Commercial systems available for multiwell planar patchclamp include patchxpress 7000a. Development of planar patch clamp technology and its application in. Robotic multiwell planar patchclamp has become common in drug development and safety programs because it enables efficient and systematic testing of compounds against ion channels during voltage. Development of planar patch clamp technology and its.
In 2009 nanion was awarded the german founders award grunderpreis, and the innovation prize step award. Dissolve 4 mg of agar in 100 ml of 3m kcl solution by boiling and transfer to 10cm dishes. One of these breakthroughs is the development of automated planar patchclamp. Nanion develops and markets instrumentation patch clamp devices, related consumables for instance, patch plates, and services for automated patch clamping. A unique feature of nanion s automated patch clamp platforms is the possibility to exchange intracellular components. A custom made holder for the microscope addon was built in home and used in combination with an. Planar patch clamp approach to characterize ionic currents. 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. For many years now manual patchclamping has been regarded as one of the. Neobiosystems automated patch clamping automate scientific. Nanions syncropatch 96 convinces big pharma automated. Planar patch clamp is a novel method developed for high throughput electrophysiology. 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.
Get a free quote for a complete electrophysiology rig setup, or just the pieces you need. This aspect is exploited with the internal perfusion system for the portapatch. We have investigated multiwell planar patchclamp for studies of wholecell macroscopic currents but the reader should be aware that single channel events are detectable in planar cellattached patch clamp recordings8 and it might be possible to generate a configuration similar to the insideout patch if the wholecell membrane is permeabilised. The portapatch is a onecellatatime system, which is surprisingly small for a complete patch clamp rig. Modelled on the success of the porta patch, the porta patch mini is also designed to be an easytouse basic research patch clamp instrument, perfect for academic laboratories that teach. Using automated patch clamp electrophysiology platforms in. Crosssite and crossplatform variability of automated.
With the ezgseal pressure controller, patch clamping becomes as easy as a click. 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. Contact is then made with the cell, and through application of negative pressure, an electrical seal forms, in the order of g. Traditional patch clamp has a very low throughput which has been proven to be a. How to access their electrophysiology by using the surfe2r technology and planar patch clamp surfe. With the latest developments in the traditional patch techniques such as wholecell and single channel as well as perforated patch, fast. For patchliner experiments, a patchliner octo nanion.
An introductory guide to patch clamp electrophysiology is a concise introduction to the basic principles and practical applications of this important technique. Launch of the dynamic clamp unit for the patchliner the. 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. Please use one of the following formats to cite this article in your essay, paper or report. Nanion vesicle prep pro bilayer apparatus automate. Nanion s planar patch clamp chips for the portapatch not only allow for recordings of cell membranes but also of artificial bilayers. Nanion combines bio and microtechnology in a company serving the life sciences industry by offering products and services which will dramatically increase the speed and efficiency of the drug discovery process in an important segment of the pharmaceutical market.
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 porta patch enables fast fluid exchange on the chip, and is suitable for both voltage gated and ligandgated ion channels. Most platforms utilize borosilicate glass configured as a pipette, however more recently planar patch clamp chips have been developed that require less technical expertise. As in conventional patch clamp, agar bridges are used for the reference electrode. The portapatch is routinely provided with an axonmdc multiclamp amplifier or an epc10 amplifier from heka together with a computer and the software packages. Nanion portapatch planar patch clamping automate scientific. However, many of the protocols and notes given in this chapter can be applied to other automated patchclamp platforms. The availability of standardized cell lines expressing ion channels of interest lead to the development of diverse automated patch clamp apc systems with highthroughput capabilities. For the patch clamp recordings, planar borosilicate glass patch clamp chips were used18 in a 384microtiterplate format. Planar patch clamping, as the name implies, is a twodimensional or planar variation on. The company offers wide range of patch clamp devices, techniques, and electro physiological products. Robotic multiwell planar patchclamp for native and.
There is a book patch clamp analysis advanced techniques, chapter 14 planar patch clamping, which i. Progress in automating patch clamp cellular physiology luca a. Specialized tools for electrophysiology and cell biology research. Ionworks quattro and ionworks barracuda, molecular devices, llc.
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. Ampio pharmaceuticals announces agreement to acquire key delivery technology. Instead of positioning a pipette on an adherent cell, cell suspension is pipetted on a chip containing a microstructured aperture. Patchliner patchliner versatile and sophisticated the patchliner is a fully automated planar patch clamp instrument recording from up to 8 cells simultaneously. Perfusion chamber i this perfusion chamber is designed for oocyte twoelectrode voltage clamp and patch clamp. Typically, cho and hek tumorigenic cell lines overexpressing single.
The patch clamp technique is widely used for recording the activity of ion channels in single cells and lipid bilayers. Nanion technologies and data analysis datacontrol 384, nanion technologies. The syncropatch 384pe is the first high quality, automated patch clamp system with the potential to bridge the gap between primary and secondary ion channel drug screening. Our planar lipid bilayers are obtained from guvs prepared with the nanion vesicle prep pro setup. Highthroughput electrophysiology with planar patch clamping. Experiments are performed on a single cell at a time, whereby whole dose response curves can be. Imagine taping a conversation, only to have the recorder make so. A single cell is then positioned on the hole by suction and a tight connection gigaseal is formed. In addition, in planar patch clamp it is also possible to use internal agar bridges. Does anyone use the automated patch clamp, the porta patch. Npc16 patchliner and syncropatch 96 nanion technologies gmbh.
This protocol assumes basic knowledge of patch clamp methods. These assets have been moved to our warehouse in canton, ma and are available for inspection. Nanion offers analysis instruments for ion channel analysis. And it contains all equipment required for experiments in only one box, among this the liquid handling components, the electrophysiological hardware and even the computer. 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. In 2009, nanion lauched its high throughput, highquality automated patch clamp system, the syncropatch 96. During a recent demonstration of the porta patch in the department of pharmacology at the university of oxford, uk, we successfully demonstrated the possibility of. Cell suspensions for apc devices utilizing planar patch clamp chips should be homogeneous milligan et al.
Besides the great equipment, nanion, with their widespread. Patch clamp measurements were performed by an automated multiwell planar patch clamp system the syncropatch 384 patch engine pe nanion technologies, munich, germany fig. 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. Novel screening techniques for ion channel targeting drugs. A comparison of the performance and application differences between manual and automated patch clamp techniques.
The system uses nanion s planar patch clamp chips and generates high quality data. 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. Nanion s team has developed and successfully established four generations of automated patch clamp instruments for sophisticated and high throughput applications in. The patch clamp technique wikipedia1 is widely used for the analysis of ion channels, which play an important role in many. Nanion synchropatch 384 for automated, highthroughput. Each system is built for your specific application so you can start using it right out of the box. A mpc involves the use of a glass pipette that is manoeuvred to an adherent cell approach. These cells were investigated by action potential analysis with manual and automatic planar patch clamp technologies, as well as by recording extracellular field potentials using a microelectrode array system. A successful robotic approach of the patch clamp technique is based on planar patch clamp chips where a glass pipette, as used in conventional patch clamping, is replaced by a thin planar. The planar lipid bilayers can be prepared on the chip either by the painting method or by guv giant unilammellar. Data output and compound information is compatible with most database formats. Experiments are performed on a single cell at a time, whereby whole dose response curves can be obtained from a single cell.
It has a much smaller foot print compared to a conventional rig. Nanion technologies gmbh manufactures medical appratus. The technique can be applied to a wide variety of cells, but is especially useful in the study of excitable cells such as neurons, cardiomyocytes, muscle fibers. Vanderbilt requests funds for the purchase of a nanion synchropatch 384 automated. Automated planar patch clamp systems are widely used in drug evaluation studies because of their ability to provide accurate, reliable, and reproducible data in a highthroughput manner. In these automatic devices like the planar population array patch clamp, papc, the micropipettes are replaced by pores in planar supports. Automated patch clamp meets highthroughput screening. The patch clamp goes planar the scientist magazine. With conventional patch clamp this is difficult because of the tiny dimensions of the pipette tip, the long tapered shaft and, thus, the.
Welcome to nanion technologies committed to high performance electrophysiology. Molecular devices, axon instrument, sophion bioscience, cytocentrics and nanion technologies have already launched their planar patch. An optional perfusion system and temperature control device is available. The syncropatch 384 pe was a patch clamp module that was integrated into a liquid handling robot owning a 384 pipetting head, biomek fx. Usa and syncropatch data analysis package, nanion technologies. We have investigated multiwell planar patch clamp for studies of wholecell macroscopic currents but the reader should be aware that single channel events are detectable in planar cellattached patch clamp recordings8 and it might be possible to generate a configuration similar to the insideout patch if the wholecell membrane is permeabilised. Automated temperaturecontrolled herg test on the patchliner. The patchliner is an automated patch clamp robot for medium thoughput with ultimate flexibility for assay design. Nanion s planar patch clamp chips offer the unique possibility to perfuse the intracellular side of the membrane.
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