This guide provides information about the patch clamp rig. Portapatch planar patch clamping the portapatch is a onecellatatime system, which is surprisingly small for a complete patch clamp rig. Using conventional patch clamp, a single trained electrophysiologist is able to produce 010 points of data per day because of the constraints of cell viability, the sequential patch of cells and the temporal nature of many of the protocols. Nanion portapatch applications automate scientific. Both of these can be useful in compound mechanism of action studies. Progress in automating patch clamp cellular physiology luca. The flexcyte 96 brings in vitro contraction force measurements of cardiac cells to a new dimension. Instead of positioning a pipette on an adherent cell, cell suspension is pipetted on a chip containing a microstructured aperture.
Nanions planar patch clamp chips offer the unique possibility to perfuse the intracellular side of the membrane. 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. 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. However, many of the protocols and notes given in this chapter can be applied to other automated patchclamp platforms. The planar lipid bilayers can be prepared on the chip either by the painting method or by guv giant unilammellar. Oct 24, 2012 introduction the patch clamp technique is a laboratory technique in electrophysiology that allows the study of single or multiple ion channels in cells. Robotic multiwell planar patchclamp for native and primary.
Traditional patch clamp has a very low throughput which has been proven to be a. Select any or all components you need for your custom electrophysiology rig. 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. They move your patch pipette with nanometer precision. Since the volume of perfusion chambers oocyte well is only 4 microliter, the solution change is almost instantaneous.
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. Experiments are performed on a single cell at a time, whereby whole dose response curves can be obtained from a single cell. Robotic multiwell planar patchclamp for native and. A single ion channel conducts around 10 million ions per second. Patch clamp manipulators are highly precise instruments. Our ultrasmall, flexible, nanowire probes could be a very powerful tool as they can measure intracellular signals with amplitudes comparable with those measured with patch clamp techniques. An introductory guide to patch clamp electrophysiology is a concise introduction to the basic principles and practical applications of this important technique. Automated patch clamp apc technology was first developed at the turn of the millennium. Most platforms utilize borosilicate glass configured as a pipette, however more recently planar patch clamp chips have been developed that. Molecular devices model cells, cables, odd bits automate. Fully automated dynamic patch clamp for the patchliner.
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. Planar patch clamps planar patch clamping, as the name implies, is a twodimensional or planar variation on traditional patch clamping. It offers fast and easy access to high quality patch clamp data with only minimal training. The estimates i give are based on traditional patch clamp rigs. Webinar patch clamp made easy, fast track to excellence. Miniaturized patch clamp system supporting gigaseal recordings from one cell at a time. 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. Oocyte is confined in a cone shaped well to a fixed position. Heat activation, action potentials, and high throughput in ion channel screening.
And the quality of the recordings rivals that of a conventional patch clamp rig. A new technique for multiple reuse of planar patch clamp. The porta patch mini contains a miniature digital amplifier integrated into the system creating the smallest and lightest patch clamp rig on the market. Sakmann and neher develop the patch clamp technique in 1970s and early 1980s. Due to their planar chip design not only is a higher throughput achieved, but new applications have also become possible. Crosssite and crossplatform variability of automated. Planar patch clamp combined with atomic force microscope afm. Just drop one or two oocytes into the included perfusion chamber and start recording. With conventional patch clamp this is difficult because of the tiny dimensions of the pipette tip, the long tapered shaft and, thus, the. Planar patchclamp biophysics and molecular materials lmu. Using automated patch clamp electrophysiology platforms in.
Get a free quote for a complete electrophysiology rig setup, or just the pieces you need. It is an important e electrophysiology technique with a broad range of applications. 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. 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 branch of physiology concerned with determining the basic mechanisms by which electric currents are generated within living organisms. Nanion vesicle prep pro bilayer apparatus automate. A unique feature of nanion s automated patch clamp platforms is the possibility to exchange intracellular components. The system uses nanions planar patch clamp chips and generates high quality data. Highly parallel planar patch clamp for ion channel screening. Perfusion chamber i this perfusion chamber is designed for oocyte twoelectrode voltage clamp and patch clamp. The data quality and reproducibility of pharmacological experiments is consistent between the automated systems and a manual patch clamp rig or semiautomated patch clamp system like the porta patch as shown for current clamp recordings from pluripotent stem cellderived cardiomyocytes. However, patch clamping is a laborious process requiring a skilled experimenter and is, therefore, not compatible with the high throughput needed in drug development.
This project focuses on the simultaneous measurements of ionchannels using ligand tethered afm tips combined with a planar patch clamp device. Novel screening techniques for ion channel targeting drugs. It is thus of special interest in the research of excitable cells such as neurons, cardiomyocytes and muscle fibers. 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. The highthroughput system used here is a planar patch clamp screening. Nanion s planar patch clamp chips offer the unique possibility to perfuse the intracellular side of the membrane.
Unfortunately, electrode instability due to the much larger compound currents, the struggle to maintain continuous voltage clamp, and the inability to have the. Comparison of a typical pipettebased patch clamp rig on the top with a planar patch clamp rig on the bottom. The patchliner is a fully automated planar patch clamp instrument recording from. Instrumentation based on the planar patch clamp principle is made available by nanion technologies gmbh. Microchip technology for automated and parallel patch. The porta patch is a miniaturized patch clamp system supporting gigaseal recordings from one cell at a time. 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. There is a book patch clamp analysis advanced techniques, chapter 14 planar patch clamping, which i. Nanion portapatch planar patch clamping automate scientific. Tedious manual patchclamp rigs have given way to automated planar patchclamp systems with sufficient throughput to play a role in the drugdevelopment process. The portapatch enables fast fluid exchange on the chip, and is suitable for both voltage gated and ligandgated ion channels. Miniaturized patch clamp system supporting gigaseal recordings from one.
This aspect is exploited with the internal perfusion system for the portapatch. 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. Patch clamp definition of patch clamp by medical dictionary. Progress in automating patch clamp cellular physiology. 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. In a collaboration with nanion, the planar patch clamp technology turned out to be a perfect platform for afm experiments with nonadherent cells. Virtual tour of whole cell patch clamp electrophysiology at the allen institute for brain science. Our planar lipid bilayers are obtained from guvs prepared with the nanion vesicle prep pro setup. 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. Nanion offers analysis instruments for ion channel analysis, as patch clamp, impedance and bilayer recordings, used for drug development as cardiac safety and basic research. The system uses nanion s planar patch clamp chips and generates high quality data. Highthroughput electrophysiology with planar patch clamping.
Neobiosystems automated patch clamping automate scientific. 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. How much it will cost on the average to obtain and set up patch clamp technology. One of these breakthroughs is the development of automated planar patchclamp technology. To measure whats happening in or on a single, living cell, scientists use a technique called the patch clamp which requires an extremely fine pipet held tightly against the cell membrane.
Planar patch clamp article about planar patch clamp by the. Received the nobel prize for this high scientificwork in1991. Two new thirdgeneration automated patch clamp apc systems capable of high. 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. A recent explosion in the number and different approaches taken to planar array patch clamp of mammalian cells has. Portapatch planar patch clamping the porta patch is a onecellatatime system, which is surprisingly small for a complete patch clamp rig. 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. An optional perfusion system and temperature control device is available. Development of planar patch clamp technology and its. 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. 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. An introductory guide to patch clamp electrophysiology at. This webinar shows applications that go way beyond possibilities of conventional patch clamping, where the porta patch facilitates completely novel scientific directions. The system uses nanions planar patch clamp chips and generates high.
The patch clamp technique is widely used for recording the activity of ion channels in single cells and lipid bilayers. Molecular devices, axon instrument, sophion bioscience, cytocentrics and nanion technologies have already launched their planar patch clamp systems separately. The increased throughput it afforded promised a new paradigm in ion channel recordings, offering the potential to overcome the time. The patchliner is an automated patch clamp robot for medium thoughput with ultimate flexibility for assay design.
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. With its vast experimental freedom and gigaseal data quality, the patchliner is one of the most versatile patch clamp instruments on the market. Thus, the introduction of automated planar patch could increase the rate of data acquisition 100fold. The here utilized medium throughput apc device is a planar patch clamp system capable of recording up to eight cells simultaneously. It is a research grade instrument allowing for whole cell as well as single channel recordings with great experimental versatility. 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. Does anyone use the automated patch clamp, the portapatch. 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. Each system is built for your specific application so you can start using it right out of the box. Solutions for action potential ap recording in the conventional patch clamp rig. Not only a powerful research tool but also ideal for educational purposes and quick tests of cells and ion channels. 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. Patch clamp recording is an extremely useful technique for investigating the biophysical properties of the ion channels that control neuronal activation. 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.
Faraday cage heka table top faraday cage tmc benchtop faraday cage the cage around your patch clamp rig shields noise from interference sources in order to get a perfect signaltonoise ratio. Fully automated planar patch clamp instrument recording from up to 8 cells simultaneously. In addition, in planar patch clamp it is also possible to use internal agar bridges. Automated patch clamping finally achieves high throughput. 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. Designed for seamless integration into processautomated drug screening environments, the patch engine is equipped with 384 patch clamp amplifiers and an advanced 384 channel liquid handling robot. How much it will cost on the average to obtain and set up. The procedure involves pressing a glass micropipette against a cell in order to isolate a small patch of membrane that contains one or more ion channels. 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. Digidata daq and pclamp software for the ultimate oocyte recording rig. Welcome to nanion technologies committed to high performance electrophysiology. Pc rig and turns it into an easytouse high throughput electrophysiology system. Automated platforms have demonstrated the ability to.
Traditionally, single channel patch clamp measurements. Atomic force microscopy and patch clamp are two incompatible techniques as long as patch clamp will be routinely done using glass pipettes. Planar patch clamp is a novel method developed for high throughput electrophysiology. A single cell is then positioned on the hole by suction and a tight connection gigaseal is formed. Planar patch clamp approach to characterize ionic currents. 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. Patch clamp electrophysiology steps up a gear european. Frontiers stateoftheart automated patch clamp devices. May 20, 2005 the porta patch npc1 from nanion is a neat benchtop instrument that has most of the functionality of a conventional patch clamp rig, including cell attached single channel recording and drug addition to the internal face of the plasma membrane.
Population patch clamp significantly improved upon the success rate, compound throughput, and welltowell variability of singlehole automated planar patch clamp john et al. Nanion s planar patch clamp chips for the porta patch not only allow for recordings of cell membranes but also of artificial bilayers. Dissolve 4 mg of agar in 100 ml of 3m kcl solution by boiling and transfer to 10cm dishes. As in conventional patch clamp, agar bridges are used for the reference electrode. Features such as temperature control and recordings in the current clamp mode. The impact of automated patch clamping apc on ion channel screening is now. It has a much smaller foot print than the conventional rig. 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. Adhesion force measurements on cells requires immobilization of the interacting partners to the substrayte and to the force sensor respectively. By carefully heating and pulling a small glass or quartz capillary tube, a very fine pipet can be formed.
The patch clamp technique allows the investigation of a small set or even single ion channels. The patchliner is a fully automated planar patch clamp instrument recording from up to 8 cells simultaneously. This guide provides information about the patchclamp rig. 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.