SURFE²R 96SE - 高通量转运体研究设备

与 SURFE²R N1一样，SURFE²R 96SE基于固体支撑膜（SSM）电生理技术，解决转运体与离子泵电流过小的问题，同时将通量提高到平行进行96个记录。只需要将转运体样品加到96孔记录板镀金电极上。并且为了进一步减少用户的工作量，所有电极预处理步骤都由SURFE²R 96SE自动完成。并且可以加任何底物都可以加入激活转运体，且会记录相应的转运或绑定电流。所有的以上步骤，从制备样品、测定到数据分析都是自动进行的。

SURFE²R 96SE的特点

• 通过内置的96通道低噪音放大器平行进行96孔记录
• 每天可获取10,000 个数据点
• 整合 CyBio^® Felix 小型移液平台
• 自动化的实验流程，包括自动制备样品
• 自动数据分析与导出，例如测定EC₅₀等
• 拥有8个放置部同溶液的位点 (支持1孔板到96孔板)

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EC_50、 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。SURFE²R 96SE相对于SURFE²R N1 拥有更高的通量。更适用于诸如待测物或转运体筛选项目。SURFE²R 96SE也适用于需要较高通量的基础研究。其数据获取速度相较于 SURFE²R N1快了将近100倍。.

更多信息:

SURFE²R 96SE 设备与软件

The SURFE²R 96SE 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 N1 as an easy-to-learn all-in-one device for universities and basic research and the high-throughput device SURFE²R 96SE.

The SURFE²R 96SE is integrated into the CyBio^® Felix liquid handling platform. Nanion Technologies provides the electrophysiological hardware and the computer running a software package for data recording and analysis. The device can not only be used for electrophysiological measurements, but also for sample preparation and pipetting purposes. The device can measure 10,000 data points a day in a fully automated manner. In contrast to the SURFE²R N1, the SURFE²R 96SE also comes with a dedicated data analysis software which allows for automated analysis of the massive amount of data you generate each day.

The SURFE²R 96SE uses sensors in a standard 96-well plate format. Membranes containing the protein of interest are adsorbed onto these sensors. The measurement is initiated by a substrate concentration jump. The solution exchange is controlled by the liquid handling platform CyBio^® Felix. The pipetting head contains 96 pipettes which load solutions from solution positions and inject them to the sensor plate. All 96 sensors of the sensor plate are measured simultaneously, but different solutions or transporters can be used for each measurement.

The sensor plate is enclosed by a Faraday cage and connected to a 96-channel amplifier as well as 96 gold pins which work as reference electrodes. The CyBio^® Felix holds up to 8 well plates for 8 to 8 x 96 different solutions which allows up to 7 sequential measurements using different conditions in each automated workflow. The CyBio^® pipettor allows the measurement of fast kinetics with low solution consumption, showing a time resolution of down to 5 ms by using only 50 µl of compound solution per measurement.

软件包

Our software department developed a software package exclusively for the SURFE²R 96SE.

• The SurfControl works as the recording software and displays the data in real-time
• DataControl works as the analysis software and can be configured to automate your data analysis
• In addition, the CyBio Composer plus pre-installed workflows operate the liquid handling platform and
communicates with the recording software

耗材

SURFE²R 96SE 芯片板

SURFE2R 96SE Sensor Plate

The SURFE²R 96SE sensor plate is a proprietary innovative product by Nanion Technologies, developed for the SURFE²R 96SE. It is produced by an external partner, quality-assured in-house at Nanion headquarters and shipped from Munich to our international customers.

Material

The core structure of SURFE²R 96SE sensor plates are gold-coated sensors with 3 mm diameter. This structure is incorporated into a 96 well plate for easy handling. The sensor plate is enclosed by a Faraday cage and connected to a 96-channel amplifier during the experiment.

Available plate type

"SURFE²R 96SE Sensor Plate": 96-well recording plate for the SURFE²R 96SE (Order # 182001)

验证过的实验对象:基于 SSM的电生理技术

用户评价与案例分析

Analytik Jena - Maria Barthmes about the Cooperation on the SURFE²R Technology

Icon N1 ““We have tested several liquid-handling platforms. CyBio FeliX was the best choice for our application.”

Dr. Maria Barthmes, Product Manager SURFE²R 96SE
Nanion, Munich

Download the full case study here:

Icon N1 SURFE²R Technology Case Study (2020) (1.4 MB)
Transporter Protein Research Redefined

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Dr. Thomas Seeger & Karin V. Niessen - Statement about the SURFE²R Technology

Icon N1 “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 and Co-Authors - Interview about the SURFE²R Technology

Icon N1 ““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:

Icon N1 SURFE²R Technology Customer Case Study (2020) (1.3 MB)
Nature Scientific Reports: Characterization of the Na⁺/H⁺ exchanger NhaB on the SURFE²R N1

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Prof. Dr. Randy Stockbridge - Interview about the SURFE²R Technology

Icon N1 “We wanted to characterize the electrophysiological characteristics of our new family of transporters to find out properties such as K_m and V_max 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:

Icon N1 SURFE²R Technology Customer Case Study (2019) (1.8 MB)
Replacing Radiolabeling Techniques with the SURFE²R N1

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数据与应用

NCX1 - High Throughput Screening on the SURFE2R 96SE

Icon N1 SURFE²R 96SE data and applications:
Cells were kindly provided by Charles River.

NCX1 over-expressed in HEK cells was recorded on the SURFE²R 96SE. A membrane preparation of HEK cells expressing NCX1 was used on the SURFE²R 96SE. An activating buffer containing 100 μM Ca²⁺ was used to activate NCX1 (black and dark blue traces) after a negative control (0 Ca²⁺; light blue trace). NCX1 was activated three times after which a pharmacological experiment can be started to test for inhibitors. Wells are color-coded so that the user can get an overview of the succes of the experiment at a glance. Green wells depict resistance >500 MΩ, blue depicts wells have a resistance between 100 and 500 MΩ.

EAAT3 - Screening of 6 inhibitors

Icon 96SE SURFE²R 96SE data and applications:

Screening of 6 inhibitors for EAAT3. After negative control and full activation of the transporter, different concentrations of inhibitor were used. At the end the full activation was repeated.

Organic Cation Transporter 2 (OCT2) - Application of Choline, TEA and Methformin

Icon 96SE SURFE²R 96SE data and applications:

OCT2 transient currents are shown before and after the application of Choline, TEA and Methformin. A concentration-dependent activation of OCT2 could be obtained by all of the three compounds. Membranes without OCT2 (control membrane) were used as a negative control.

PepT1 - Pharmacology of Losartan

SURFE²R N96 (a predecessor model of SURFE²R S96) data and applications:

Transport by PepT1 is inhibited by the kompetitive antagonist Losartan. When activated with 20 mM Gly-Gly, the IC₅₀ of Losartan was 80.4 µM. Data were analysed with DataControl - the Analysis software tool for recordings on the SURFE²R N96 and SyncroPatch 96/ 384PE.

PepT1 - Evoked signal via Glycylglycin

SURFE²R N96 (a predecessor model of SURFE²R 96SE) data and applications:

Graphical user interface of screening software used on the SURFE²R N96. Traces of PepT1 recorded from membrane preparations of PepT1 overexpressing HEK293 cells are presented. The PepT1 signal is evoked by 20mM glycylglycin.

网络研讨会与影像

2019 - SURFE²R as a service - closing the gap between technology and application

Icon 96SE SURFE²R 96SE Oral Presentation

Presenter:
Dr. Ralf Schwandner, CEO of Assay.Works GmbH, Germany
Assay.Works offers assay and screening services using the SURFE²R 96SE. Find out more here.

2018 - Electrophysiological Characterization of Sugar Transporters using SSM-based Electrophysiology

Icon 96SE SURFE²R 96SE and Icon N1 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 - Pharmacological characterization of an amino acid transporter and his bacterial homologue – a case study using the SURFE²R 96SE technology

Icon 96SE SURFE²R 96SE Oral Presentation Video

Presenter:
Dr. Thomas Licher, Head of in vitro Biology & High-Throughput Chemistry, Sanofi, IDD Frankfurt, Germany
Thomas made his PhD at the Center for Molecular Neurobiology in the group of Prof. Dr. Pongs on an HTS-technology for ion channel-modulating substances in 2002. He started to work for Sanofi-Aventis, later Sanofi as Post-doc, lab head and group leader in the field for Cardiac Safety and Lead Identification Technologies. Today, Thomas is Head of in vitro Biology & High-Throughput Chemistry at Sanofi in Frankfurt, Germany.