SURFE²R N1 - 转运体工作站
与传统膜片钳设备相比，SURFE²R (表面电荷事件读取器，surface electrogenic event reader)是专用于测量带电转运体 (同向转运体、交换体与单向转运体)与离子泵的技术。 通常这些蛋白相对与离子通道的转运率较低。 SURFE²R技术通过更大的芯片面积从而同时记录最多达109 个转运体从而获取最佳的信噪比。SURFE²R N1是适用于基础研究与大学教学的设备。如果需要高通量， SURFE²R 96SE 可提供最多平行96芯片的转运体记录。
SURFE²R N1的特点
• 支持最多52种不同溶液的自动记录
• 每天可记录150数据点
• 移液设备、电生理设备与电脑整合的一体化设备
• 易学易用，适用于教学与科研
基于SSM的电生理技术特点
• 已在超过100 目标上通过验证，发表了超过 100篇同行评审过的文献
• 可测量转运体、离子泵与配体门控通道
• 也可测量电中性交换体、糖结合蛋白
• 可使用从细胞提纯的生物膜与囊泡或蛋白脂质体
• 非标记电学测量手段
• 每孔仅需要0.1 – 1 µg蛋白样品，并足够进行最多100次实验
• 实时数据与5 ms的高时间分辨率，非单点读取
• 相对膜片钳更高信号放大率
• 可完成快速绑定动力学测量
• 可完成EC50、 IC50、转运速度常数、转运体亚型比较...
基于固体支撑膜技术（solid-supported membrane，SSM) 的电生理技术建立于90年代末，关于SSM电生理技术的详细资料请参考 技术细节。 可重复使用的芯片包含一个镀金电极，在上面可以通过一次快速移液形成SSM，然后将包含转运体的样品加到SSM表面上。任何包含目标蛋白的膜制成品都可以用来测量，包括细胞破坏后形成的膜囊泡或重建提纯的蛋白形成的蛋白脂质体。这些样品可以冷冻保存数月甚至数年，因此不需要细胞培养室。
基于SSM的电生理技术的关键是通过溶液交换来施加底物或者配体来激活转运体，然后电荷的迁移可以被检测出来并被分析以得出需要的数据。 由于SSM的高稳定性，同一个样品可以进行上百个实验，所以可以测定诸如EC50 或IC50等参数 。拥有目前最先进的溶液交换的时间分辨率，SURFE²R N1不但可以测量慢的转运过程，也可以分析快速的结合反应以及确定其反应速度常数。
SURFE²R N1易学易用，可以在一天时间内学会并掌握。该设备通过机械臂自动进行实验，极大的减少了操作人员的工作量。SURFE²R N1在一个设备里包含了实验所需的所有设备，包括溶液处理元件、电生理硬件和电脑主机。
更多信息:
SURFE²R N1设备与软件
The SURFE2R N1 设备
The SURFE2R (surface electrogenic event reader) technology is the only available commercial solution for SSM-based electrophysiology on the market. Nanion Technologies developed the SURFE2R 96SE, a high-throughput system mainly used in pharmaceutical industry for drug screening purposes and the SURFE2R N1 designed for basic research.
The SURFE2R N1 device contains electrophysiological hardware, liquid handling components and the computer running Windows, plus the data recording and analysis software SurfControl. It’s an easy-to-learn, all-in-one robotic workstation which can measure 150 data points a day in a fully automated manner.
The SURFE2R N1 works with reusable sensor chips on which the membranes containing the protein of interest are adsorbed. The measurement is initiated by a substrate concentration jump. The solution exchange is controlled by the Ionjet, a robot-controlled pipette which loads solution from storage containers and injects them to the sensor enclosed by a Faraday cage. The Ionjet allows the measurement of fast kinetics with low solution consumption. An autosampler on top of the device allows placing up to 53 solutions for automated sequential measurements. During the measurement, the transport current can be viewed, compared and analyzed within the SurfControl software. The main characteristics of the device include a low-noise amplifier and a large sensor surface which both ensure a superior signal to noise ratio.
SURFE2R N1 控制软件与自动化
The device comes with the SURFE2R N1 Control software pre-installed on the internal computer. This forms both the recording and analysis software. The functions of SurfControl include the managing and coding of workflows which represent protocols for SURFE2R assays. Workflows can contain multiple measurements, e.g. using different substrate concentrations or the comparison of transport before and after inhibition. The workflow includes parameters like duration, speed and volume of solution flow during the experiment, the number and sequence of different buffers, the number of repetitions per measurement, the incubation times between experiments and the volume used for rinsing the sensor after an experiment. During the run of one workflow no interference by the researcher is required.
Beside the design of the experiment itself, SurfControl enables the researcher to view and compare recorded traces. The graphical window allows peak detection and the calculation of the peak integrals including baseline subtraction. An additional results window is used for automated documentation of the traces including file name, values for peaks and integrals and the time of measurement. For further analysis, e.g. the fitting of kinetic parameters or the subtraction of negative control currents, the software is capable of exporting the data to the standard file format ASCII.
耗材
SURFE²R N1单通道记录芯片
The SURFE²R N1 sensor chip is a proprietary innovative product by Nanion Technologies, developed for the SURFE²R N1. It is produced by an external partner, quality-assured in-house at Nanion headquarters and shipped from Munich to our international customers.
材质
The core structure of SURFE²R N1 sensor chip is a gold-coated sensor with 3 mm diameter. This structure is incorporated into a screw cap for easy handling. The SURFE²R N1 single sensor chips can be reused after an appropriate cleaning protocol.可用的型号
- "SURFE²R N1 Single Sensor Chip" (Order # 161001)
已经验证的对象: 基于SSM的电生理
用户评价与案例分析
Dr. Arnaud Javelle - Statement about the SURFE²R N1 Device
“Using the SURFE2R N1 we have recently obtained high quality data in a very short period of time. We have developed an assay to measure the activity of ammonium transporters from the Amt protein family. There has been considerable controversy over the mechanism of ammonium transport by Amt proteins and the controversy was due to the lack of quantitative kinetic data characterizing the activity of the proteins at the single channel level. The SSM technologies allows to overcome this hurdle and we are now capable of answering very challenging functional questions concerning the mechanisms and the energetics of these transporters."
Dr. Arnaud Javelle, Chancellor's fellow, Strathclyde Institute of Pharmacy and Biomedical Sciences
Dr. Thomas Seeger & Karin V. Niessen - Statement about the SURFE²R Technology
“The SURFE²R makes extremely challenging electrophysiological targets accessible to robust routine analyses. For example, we have developed an assay for investigating nicotinic acetylcholine receptor ion channels, using membranes from the Pacific electric ray, Torpedo California. In this way, we overcome many of the well-known difficulties associated with this ion channel, which allows us to efficiently obtain information regarding compound pharmacology. Obtained pharmacology values match those of patch clamp recordings exceptionally well. We find that the SURFE²R is an excellent platform for characterization of electrogenic processes in isolated membranes.”
Karin V. Niessen, Dr. Thomas Seeger
Bundeswehr Institute of Pharmacology and Toxicology, Munich, Germany
Prof. Dr. Klaus Fendler - Statement about the SURFE²R N1 Device
“SSM-based electrophysiology is a well-established technique developed in our lab in the early 1990s for the investigation of electrogenic membrane transporters. It works with a wide range of samples from mammalian cell membranes over microbial membrane vesicles to proteoliposomes. SSM-based electrophysiology is especially useful in cases where conventional electrophysiology cannot be applied, e.g. for transporters residing in intracellular membranes or plasma membranes of small microbial cells. The SURFE2R N1 is a commercial instrument for SSM based electrophysiology which facilitates basic research in the field of membrane transport.“
Prof. Dr. Klaus Fendler
Max-Planck Institute of Biophysics, Frankfurt, Germany
Prof. Dr. Klaus Fendler and Co-Authors - Interview about the SURFE²R Technology
““SSM-based electrophysiology is a suitable technique to study bacterial ion-translocating membrane- systems which are otherwise difficult to approach using conventional electrophysiological techniques, like patch clamp.
Additionally, the mechanical stability of SSMs facilitates the transport activation through fast solution exchange, a condition that is technically challenging in a setup of a bilayer separating two solution-containing compartments.“
Dr. Miyer Fabián Patiño-Ruiz,
Faculty of Science and Engineering, University of Groningen, the Netherlands
Dr. Andre Bazzone, Application Scientist for the SURFE²R product family interviewed the authors of the publication:
"Mutation of two key aspartate residues alters stoichiometry of the NhaB Na+/H+ exchanger from Klebsiella pneumoniae"
Published 2019 in Nature Scientific Reports.
Read the publication here and download the full case study here:
SURFE²R Technology Customer Case Study (2020) 
(1.3 MB)
Nature Scientific Reports: Characterization of the Na+/H+ exchanger NhaB on the SURFE²R N1
Prof. Dr. Randy Stockbridge - Interview about the SURFE²R Technology
“We wanted to characterize the electrophysiological characteristics of our new family of transporters to find out properties such as Km and Vmax of the substrate, and ideally to screen a set of substrates for transporter activity. However, given our difficulties in developing a transport assay, I was pessimistic that we were going to be able to move beyond radiolabeled substrates.
Contrary to my expectations, electrophysiology with the SURFE²R N1 platform worked right away. We saw very robust currents in our pilot experiments during the initial training session on the instrument.“
Prof. Dr. Randy Stockbridge, Assistant Professor
University of Michigan, USA
Download the full case study here:
SURFE²R Technology Customer Case Study (2019) (1.8 MB)
Replacing Radiolabeling Techniques with the SURFE²R N1
数据与文献
NCX1 - Block by KB-R7943
SURFE2R N1 data and applications:
Cardiac NCX1 expressed in iCell® Cardiomyocytes2, Cor.4U® Cardiomyocytes or HEK cells was blocked by increasing concentrations of KB-R7943.
NCX1 - Block by SEA0400
SURFE2R N1 data and applications:
Cardiac NCX1 expressed in iCell® Cardiomyocytes2 or HEK cells was blocked by increasing concentrations of SEA0400, a specific blocker of NCX.
NCX1 - Current traces recorded from Cor.4U Cardiomyocytes, iCell Cardiomyocytes and HEK cells
SURFE2R N1 data and applications:
Cardiac NCX1 was activated using 30 μM Ca2+ on the SURFE2R N1. NCX1 was recorded from Cor.4U® Cardiomyocytes, iCell® Cardiomyocytes2 or overexpressed in HEK cells. The calcium affinity was determined for NCX1 recorded from iCell® Cardiomyocytes2 and HEK cells, shown on the right.
AmtB - Activation by ammonium
SURFE2R N1 data and applications:
Purified AmtB from E.Coli was incorporated into liposomes and used on the SURFE2R N1. Shown are transient currents measured after a 100 mM ammonium jump in empty liposomes (green) or proteoliposomes containing AmtB at a lipid protein ratio (LPR) of 50 (black), 10 (red) or 5 (blue). Insert: Normalized current measured in proteoliposomes containing AmtB at an LPR of 50 (black), 10 (red) or 5 (blue).
Data from Mirandela et al, 2018
AmtB - Substrate specificity
SURFE2R N1 data and applications:
Purified AmtB from E.Coli was incorporated into liposomes and used on the SURFE2R N1. AmtB substrate specificity. (A) Transient current measured after a 100 mM substrate jump. Ammonium (red), methylammonium (black), potassium (green) or sodium (purple). Potassium and sodium do not act as substrates for AmtB and methylammonium translocates but at a much reduced rate compared with ammonium. Insert: Normalised current after a 100 mM substrate jump. Ammonium (red), methylammonium (black).
Data from Mirandela et al, 2018
EAAT3 - Current Traces at different Sodium Chloride Concentrations
SURFE2R N1 data and applications:
EAAT3 current traces were recorded after application of different Sodium Chloride concentrations. The EC50 of the peak current was determined as 42.2 mM.
Organic Cation Transporter 2 (OCT2) - Pharmacology
SURFE2R N1 data and applications:
EC50 values and relative Vmax for three different substrates are shown.
The IC50 value for Choline was determined as 4.6 mM, for TEA as 0.33 mM and for Metformin as 8.6 mM.
Sodium-Potassium ATPase - Analysis
SURFE2R N1 data and applications:
The Sodium-Potassium ATPase, also known as Na+/K+ pump is responsible for the active transport of Na+ and K+ in the cells containing relatively high concentrations of K+ ions but low concentrations of Na+ ions. The Na+/K+-ATPase helps maintain resting potential, avail transport, and regulate cellular volume. Here, we demonstrate the conductance of the pump, in the presence of ATP, Na+ and K+, or in the presence of ATP and Na+.
Respiratory Chain Complex V - Action of the ATP Synthase after Application of ATP
SURFE2R N1 data and applications:
The Respiratory Chain Complex V uses the transmembrane proton gradient (produced by Complex I, III and IV) to generates ATP from ADP plus phosphate. One component of ATP synthase acts as an ion channel that provides for a proton flux back into the mitochondrial matrix. This reflux releases free energy, which is used to drive ATP synthesis, catalyzed by the other component of the complex. Experiments on the SURFE2R N1 show the action of the ATPase after application of ATP.
Respiratory Chain Complex IV - Current Traces
SURFE2R N1 data and applications:
The Respiratory Chain Complex IV, known as Cytochrom c oxidase, reduces oxygen by cunsumption of electrons from cytocrome c and transport of two protons to the intermembrans space. Here the reaction runs backwards.
Respiratory Chain Complex II and III - Current Traces
SURFE2R N1 data and applications:
Respiratory Chain Complex II is a succinate dehydrogenase. In this complex, succinate is oxidated to fumarate. The electrons are transfered to the quinone pool, the Complex III (cytochrome bc1 complex) and cytochrome c, while four protons are transported to the intermembrane space.
Respiratory Chain Complex I and III - Current Over Mitochondrial Membrane
SURFE2R N1 data and applications:
In the respiratory chain Complex I and III, oxidation of NADH leads to transport of 4 protons across the membrane. The electrons are transferred to cytochrome c. Because of the electron leakage to oxygen, both complex are the main sites of production of harmful superoxide. In these experiments using the SURFE2R N1, application of 100µM NADH results in a current over the mitochondrial membrane, where Complex I and III are expressed.
Adenine Nucleotide Translocator (ANT) - Recorded on the Surfer
SURFE2R N1 data and applications:
The Adenine Nucleotide Translocator (ANT) is located in the inner membrane of mitochondria and transports, as the name suggests, either ADP or ATP alone or against each other. Recordings were done on mitochondria inner membrane preparations from pigs heart. Example traces display the action of ANT either in the presence of ATP, in an other experiment in the presence of ADP.
PepT1 - Pharmacology of Lys[Z(NO2)]-Val
SURFE2R N1 data and applications:
Representive traces of PepT1 overexpressed in CHO cells. PepT1 (SLC15A1) is expressed mainly in intestine and kidney, it enables uptake of oligopeptides. The PepT1 signal is evoked by glycylglycin. The transport of oligopeptides can be inhibited by addition of Lys[Z(NO2)]-Val. The IC50 of Lys[Z(NO2)]-Val was 380µM.
网络研讨会与影像
2018 - Electrophysiological Characterization of Sugar Transporters using SSM-based Electrophysiology
SURFE²R 96SE and SURFE²R N1 Oral Presentation
Presenter:
Dr. Andre Bazzone, Application Scientist, Nanion Technologies GmbH, Germany
Andre is an expert in the field of SSM-based electrophysiology: He made his PhD at the Max-Planck-Institute of Biophysics in Frankfurt, Germany on the electrophysiological characterization of sugar transporters using the SSM-based electrophysiology in 2016. Right afterwards, he started as Application Scientist at Nanion Technologies and today he is an important member of the Nanion SURFE²R team.
2018 - Transporters Investigated Using the SURFE²R Instruments
SURFE²R 96SE and SURFE²R N1 Oral Presentation
Presenter:
Dr. Maria Barthmes, Product Manager SURFE²R product family, Nanion Technologies GmbH, Germany
2017 - Variations on the Coupling Theme in Secondary Active Transport
SURFE²R N1 Oral Presentation
Presenter:
Gregor Madej, University of Regensburg, Germany
下载:
应用数据
CNT1 - "Electrophysiological recordings of CNT1 (SLC28A1) activity on Nanion’s SURFE²R N1"
SURFE2R N1 application note (0.4 MB)
EAAT3 - "Electrophysiological recordings of EAAT3 (SLC1A1) activity on Nanion’s SURFE²R N1"
SURFE²R application note (0.4 MB)
GAT1 - "Electrophysiological recordings of hGAT1 (SLC6A1) activity on Nanion’s SURFE²R N1"
SURFE²R application note (0.4 MB)
Neurons - "Electrophysiological recordings of LGIC and AA transporters in iCell® GlutaNeurons"
Patchliner, 
SyncroPatch 384PE (a predecessor model of the SyncroPatch 384i) and SURFE2R N1 application note: (0.5 MB)
Cells were kindly provided by FUJIFILM Cellular Dynamics, Inc.
PepT1 - "Electrophysiological recordings of PepT1 (SLC15A1) activity on Nanion’s SURFE²R"
SURFE²R N1 application note: (0.9 MB)
产品单页
快速指南
SURFE²R N1 - Quickstart Guide "Protocols"
SURFE²R N1 Quickstart Guide "Protocols"
发表文献
2019 - Phosphatidylserine flipping by the P4-ATPase ATP8A2 is electrogenic
SURFE²R ONE (a predecessor model of SURFE²R N1) publication in PNAS (2019)
Authors:
Tadini-Buoninsegni F., Mikkelsen S.A., Mogensen L.S., Molday R.S., and Andersen J.P.
2019 - Mutation of two key aspartate residues alters stoichiometry of the NhaB Na+/H+ exchanger from Klebsiella pneumoniae
SURFE²R N1 publication in Nature Scientific Reports (2019)
Authors:
Patiño-Ruiz M., Fendler K., & Călinescu O.
2019 - Functional Reconstitution of Membrane Proteins Derived From Eukaryotic Cell-Free Systems
SURFE2R N1 and 
Orbit 16 publication in Frontiers in Pharmacology (2019)
Authors:
Dondapati S.K., Lübberding H., Zemella A., Thoring L., Wüstenhagen D.A., Kubick S.
2019 - A two-lane mechanism for selective biological ammonium transport
SURFE²R N1 pre-publication in bioRxiv (2019)
Authors:
Williamson G., Tamburrino G., Mirandela G.D., Boeckstaens M., Bage M., Pisliakov A., Ives C.M., Terras E.T., Bizior A., Hoskisson P.A., Marini A.M., Zachariae U., Javelle A.
2018 - The lipid environment determines the activity of the E. coli ammonium transporter, AmtB
SURFE²R N1 publication in Faseb J. (2018)
Authors:
Mirandela G.D., Tamburrino G., Hoskisson P.A., Zachariae U., Javelle A.
2017 - SSM-Based Electrophysiology for Transporter Research
SURFE²R N1 and SURFE²R 96SE book chapter in Methody in Enzymology
Authors:
Bazzone A., Barthmes M., Fendler K.
2017 - Insights into the mechanism of membrane pyrophosphatases by combining experiment and computer simulation
SURFE²R N1 publication in Structural Dynamics (2017)
Authors:
Shah N.R., Wilkinson C., Harborne S.P.D., Turku A., Li K.-M., Sun Y.-J., Harris S., Goldman A.
2017 - Discovery of Compounds that Positively Modulate the High Affinity Choline Transporter
SURFE²R N1 publication in Frontiers in Molecular Neuroscience (2017)
Authors:
Choudhary P., Armstrong E.J., Jorgensen C.C., Piotrowski M., Barthmes M., Torella R., Johnston S.E., Maruyama Y., Janiszewski J.S., Storer R.I., Skerratt S.E., Benn C.L.
2016 - Membrane pyrophosphatases from Thermotoga maritima and Vigna radiata suggest a conserved coupling mechanism
SURFE²R N1 publication in Nature Communications (2016)
Authors:
Li K., Wilkinson C., Kellosalo J., Tsai J., Kajander T, Jeuken L.J.C., Sun Y., Goldman A.
2016 - Electrophysiological characterization of the archaeal transporter NCX_Mj using solid supported membrane technology
SURFE²R N1 and 
Vesicle Prep Pro publication in Journal of General Physiology (2016)
Authors:
Barthmes M., Liao J., Jiang Y., Brüggemann A., Wahl-Schott C.
2014 - Anticancer Ruthenium(III) Complex KP1019 Interferes with ATP-Dependent Ca2+ Translocation by Sarco-Endoplasmic Reticulum Ca2+-ATPase (SERCA)
SURFE²R N1 publication in ChemMedChem (2014)
Authors:
Sadafi F.Z., Massai L., Bartolommei G., Moncelli M.R., Messori L., Tadini-Buoninsegni F.
2013 - Enhanced adsorption of Ca-ATPase containing vesicles on a negatively charged solid supported membrane for the investigation of membrane transporters
SURFE²R N1 publication in Langmuir (2013)
Authors:
Sacconi A., Moncelli M.R., Mergheri G., Tadini-Buoninsegni F.
2009 - Measuring Ion Channels on Solid Supported Membranes
SURFE²R ONE (a predecessor model of SURFE²R N1) publication in Biophysical Journal (2009)
Authors:
Schulz P., Dueck B., Mourot A., Hatahet L., Fendler K.
2006 - Establishment of Cell-Free Electrophysiology for Ion Transporters: Application for Pharmacological Profiling
SURFE²R ONE (a predecessor model of SURFE²R N1) publication in Journal of Biomolecular Screening (2006)
Authors:
Geibel S., Flores-Herr N., Licher T., Vollert H.
海报
2018 - Label-free analysis of Na+/Ca2+- exchanger (NCX) isolated from iPSC-derived cardiomyocytes
SURFE²R N1 and 
SURFE2R 96SE poster, Europhysiology Meeting 2018 (1.5 MB)
2018 - Transported by light: optogenetic control of NCX1
SURFE²R N1 poster, Biophysics Annual Meeting 2018 (2.2 MB)
2015 - Organellar Transporters and Ion Channels - How to access their electrophysiology by using the SURFE2R technology and Planar Patch Clamp
SURFE²R N1 and SyncroPatch 96 (a predecessor model of the SyncroPatch 384PE) and 
Port-a-Patch poster, GRC - Organellar Channels and Transporters 2015 (1.6 MB)
2017 - An emerging technique for the characterization of transport proteins: SSM-based electrophysiology
SURFE²R N1 and SURFE²R SE96 poster, 19th IUPAB / 11th EBSA congress 2017 (3.3 MB)
2015 - Functional Characterization of Procaryotic NCX by Solid Supported Membrane Technology
SURFE²R N1 poster, Gordon Research Conference 2015 (0.9 MB)
