Orbit 16 TC - Taking the Pain out of Painting
Orbit 16 TC(オービット16)は、人工脂質二重膜を用いた実験における、脂質膜の形成/膜タンパク質の再構成/単一チャネル電流の測定を簡単に行うことができます。
一度に16 chの人工脂質二重膜を迅速に自動形成し、完全同時測定を行うことで、人工脂質二重膜実験を用いた実験におけるpainting法の苦痛から研究者を解放します。
Orbit 16 TCの特徴と優位性は以下の通りです。
- 16chの人工脂質二重膜をボタン一つで自動形成/同時測定が可能
- 低ノイズ、高帯域幅での完全同時測定
- 温度コントロール可能(5~50℃)
- 専用の測定ソフトウェア同梱
- 標的チャネルタンパク質を直接再構成または、プロテオリポソームで膜融合
- イオンチャネル:電位依存性、リガンド依存性、温度依存性
- ナノポア、抗菌ペプチド、トキシンなど...
- ティスポ式 MECA 16 TC チップによる低コスト実験
Orbit 16 TC は 専用の低ノイズ16 chアンプ (Elements社 S.R.L.) を搭載し、広帯域幅での16chの脂質二分子膜の完全同時測定を行うことができます。
Orbit 16 TC バイレイヤー用プラットフォーム構成
Orbit 16 TC システムは、レコーディングステーション、内蔵型のElements社製 e16n バイレイヤーアンプ、温度制御システム、専用のワークステーション(PC)で構成されています。
詳細情報:
The intuative and easy to learn EDR4 software for the Orbit 16 TC was developed by our partner Elements S.R.L.(Italy). Further detailed information can be found at https://elements-ic.com/
The MECA 16 TC recording substrate contains an array of 16 circular microcavities in a highly inert polymer. Each cavity contains an individual integrated Ag/AgCl-microelectrode. The bilayer is automatically formed by remotely actuated painting (Ionera1-SPREAD), thus roofing the liquid-filled cavity. The bilayers can be easily and repeatedly zapped and re-formed in an automated fashion. After bilayer formation, ion channels or nanopores are reconstituted via self-insertion, proteoliposome fusion or dilution from detergent micelles.
The MECA 16 TC recording chips are produced and quality assured by our partner Ionera Technologies GmbH in Freiburg Germany and shipped from Munich to our international customers. Different types of MECA 16 chips will be available depending on the sample.
Dr. Friedrich Simmel, Professor, Systems Biophysics and Bionanotechnology, Physics Department and ZNN/WSI Dr. Stefan Howorka, Associate Professor of Organic Chemistry and Chemical Biology Event-averaged histograms (black) and overlaid current traces (blue) of parallel and simultaneous recordings on a MECA chip of monoPEG-28-mediated blockages of hemolysin nanopore(s). The current traces were recorded with a multichannel amplifier (Tecella Jet 16). Histograms were derived from the mean current levels of at least 2000 visits of blocked stated per cavity (20 kHz sample frequency). Current traces and histograms derived from recordings of αHL pores blocked by monoPEG-28 and polyPEG-1500 on an Ionera MECA chip (AxoPatch 200B, filter freq: 20kHz, digitized at 200 kHz). The data image shows parallel recordings from reconstituted alamethicin channels. See also the paper: "Alamethicin Supramolecular Organization in Lipid Membranes from 19F Solid-State NMR", Salnikov et al. (2016) Biophysical Journal 111(11): 2450-2459. Automated formation of membranes from polyoxazoline based triblock popolymers. Screenshot of a recording of Alpha-Hemolysine in a polyoxazoline based triblock copolymer membrane on the Orbit 16. Alpha-Hemolysin is capable of insertion into triblock copolymer membranes. Single channel currents of tetrameric potassium channel KcsA E71A recorded from 5 selected bilayers in parallel. Traces illustrating RYR single channel activity in the planar lipid bilayer recorded on the Orbit 16. (A) Activity after vesicle fusion and buffer exchange on cis-side. (B) 120 s after addition of 1 mM Na-ATP to the cis-side. (C) 120 s after addition of 5 µM Ryanodine to the cis-side. Screenshot of the recording window showing simultaneous and parallel assay of channel-forming activity and single-channel conductance of recombinant MspA mutant porin in a diphytanoyl phosphatidylcholine bilayer derived in 1 experimental run with the Orbit 16. Traces from a single experiment recorded in parallel from 16 lipid bilayers. Grids X: 1 s; Y: 100 pA. Addition of MspA in OPOE detergent micelles resulted in insertion of 97 pores in 12 bilayers. Screenshot of the recording window showing simultaneous and parallel PEG detection with single aHL-nanopores. Channels 1-5,7,12-14 contain a single aHL-nanopore. Channels 10 and 11 have two and Channel 9 has three aHL-nanopores. In Channels 8 and 14 single aHL-nanopores are assembled as hexamer. Channels 6 and 16 are switched off. Screenshots of a recording window of a typical Gramicidin ion channel forming activity assay on the Orbit 16.
Date: September 17. 2020, 4:00 PM CET (10:00 AM EDT) Speakers: Dr. Conrad Weichbrodt (Senior Scientist / Product Manager Orbit family; Nanion Technologies) Learn about single channel measurements in bilayer recording using the orbit instrument family - Simplifying artificial bilayer experiments: Single-molecule experiments on micro-cavity arrays Presenter: Presenter: ソフトウェア
Orbit 16 TC EDR4 Software
消耗品
MECA 16 TC chips
Available chip types
インタビュー & ケーススタディ
Prof. Dr. Friedrich Simmel - Statement about the Orbit 16
“The Orbit 16 enables us to generate high quality, single channel recordings with synthetic DNA membrane channels, which in our experience are notoriously difficult to measure. DNA pores are quite hard to functionally incorporate into lipid bilayers, but could be successfully investigated using the Orbit 16, as published in Science. The Orbit 16 offers a drastic increase in throughput since it substantially speeds up formation of bilayers and data generation by its parallel recording channels, thus providing us an easy-to-use platform for efficient and accurate research on DNA nanodevice-membrane interactions.”
Technical University of Munich, Munich, Germany
Prof. Dr. Stefan Howorka - Statement about the Orbit 16
“Within our research on the CsgG channel, Nanion’s Orbit 16 - combined with Ionera's MECA Chip technology - has substantially boosted our scientific output. The outstanding research tool is easy to handle and speeds up the parallel generation of 16 bilayers. By increasing the throughput of single-channel current recordings, it is a breakthrough in the biophysical analysis of pore forming proteins. Within approximately one week’s worth of lab time, we had the data needed for the recent paper in Nature. This would have been hard to achieve using conventional serial bilayer methods. In addition, the ease-of-use provided by the Orbit 16 shortens the learning curve for making high quality bilayer recordings. As a benefit in academia, students can now get hands-on experience with bilayer recordings also for shorter projects.“
University College London, London, UKデータ & アプリケーション
Alpha-Hemolysin - Parallel Recordings of monoPEG-28 Block
Orbit 16 data and applications:
Data courtesy of Dr. Gerhard Baaken et. al., University of Freiburg / Ionera.
Read the full paper. (Am. Chem. Soc Nano, 5(10), 8080-8088, 2011)
Alpha-Hemolysin - Block by Mono- and Poly PEGs
Orbit 16 data and applications:
Data courtesy of Dr. Gerhard Baaken et.al, University of Freiburg / Ionera.
Read the full paper: (Am. Chem. Soc Nano, 5(10), 8080-8088, 2011)
Alamethicin - Parallel recordings
Orbit 16 data and applications:
Data courtesy of Dr. Gerhard Baaken, University of Freiburg / Ionera.
Alpha-Hemolysin - Automated Formation of Membranes from Polyoxazoline based Triblock Copolymers
Orbit 16 and applications:
Data were kindly provided by Ionera.
(A) Current-voltage relationship of Alpha-Hemolysin pore in Poly(2-methyloxazoline-b-dimethylsiloxane-b-2-methyloxazoline) membrane. Average of two channels. Conditions: 25 mM Tris, 4 M KCl, pH 8.0.
(B+C) Representative recordings of Alpha-Hemolysin with PEG-28 at 40 mV and -40 mM. Conditions: 25 mM Tris, 4 M KCl, pH 8.0. Note different time scale at positive (B) and negative (C) potentials.
KcsA - Single Channel Recordings
Orbit 16 and applications:
Data were kindly provided by Ionera.
KcsA was expressed in vitro with its co-translational integration into liposoms containing asolectin lipids. The proteoliposomes were subsecuently fused with bilayer array containing POPE/POPG on the Orbit 16.
Conditions: Current traces were recorded in 20 mM MES pH 4.0 on the cis-side and 10 mM MOPS, pH 7.0 on the trans-side of the bilayer; containing 200 mM KCl symmetric solutions with membrane potential held at +150 mV.
Ryanodine Receptor - Application of Na-ATP and Ryanodine
Orbit 16 and applications:
Data were kindly provided by Ionera.
The RYR channel was reconstituted via fusion of sarcoplasmic reticulum vesicles with preformed asolectin bilayer.
Conditions: Trans-side: 53 mM Ba(OH)2, 1 mM Ca2+; Cis-side: 150 nM Ca2+, VHold: 0 mV in all cases
MspA - Mycobacterial Porin
Orbit 16 and applications:
Data were kindly provided by Ionera.
Conditions: 20 mM HEPES, 350 mM KCl, pH 7,5, rMspA final concentration 20 ng/ml; holding potential +40 mV
Alpha-Hemolysin - PEG Detection
Orbit 16 and applications:
Data were kindly provided by Ionera.
Conditons: 3 M KCl, 20 mM TRIS, pH 8, +40 mV
Gramicidin - Ion Channel Forming Antibiotic
Orbit 16 and applications:
Data were kindly provided by Ionera.
Conditions: symmetrical 0,1 HCl, +150 mV.ウェビナー & 動画
Webinars
17.09.2020 | Webinar: Electrophysiological investigation of integral membrane proteins using the Orbit mini
Orbit mini Webinar
28.06.2018 | Webinar: Artificial Lipid Bilayers in focus: Hand-held DNA-sequencing and biosensing with nanopores
Orbit mini and
Orbit 16
- Hand-held DNA-sequencing and biosensing with nanopores
27.01.2016 | Webinar: Instant bilayers - just add protein
Orbit 16 and
Orbit Mini
This webinar covers the use of the lipid bilayer platforms from Nanion: the Orbit16 and the Orbit mini for characterization of membrane proteins like ion channels, bacterial porins and biological nanopores. Both bilayer systems support high quality low noise recordings, but differ in throughput capabilities and experimental features. The Orbit16, introduced in 2012 is a device for efficient formation of 16 lipid bilayers simultaneously, allowing for parallel bilayer-reconstitution of ion channels and nanopores.
Movies: Oral Presentations and Tutorials
2018 - Simplifying artificial bilayer experiments: Single-molecule experiments on micro-cavity arrays
Orbit 16 and
Orbit mini Oral Presentation
Dr. Conrad Weichbrodt, Product Manager Orbit instrument family, Nanion Technologies GmbH, Germany
Source:
Webinar: "Artificial Lipid Bilayers in focus: Hand-held DNA-sequencing and biosensing with nanopores", June 28, 2018
2018 - Hand-held DNA-sequencing and biosensing with nanopores
Orbit 16 Oral Presentation
Prof. Dr. Stefan Howorka, University College London, Department of Chemistry
Source:
Webinar: "Artificial Lipid Bilayers in focus: Hand-held DNA-sequencing and biosensing with nanopores", June 28, 2018
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製品カタログ
論文
2022 - The Potential of Eukaryotic Cell-Free Systems as a Rapid Response to Novel Zoonotic Pathogens: Analysis of SARS-CoV-2 Viral Proteins
Orbit 16 publication in Frontiers in Bioengineering and Biotechnology (2022)
Ramm F., Dondapati S.K., Anh Trinh H.A., Wenzel D., Walter R.M., Zemella A., Stefan Kubick S.
2022 - Resolving Isomeric Posttranslational Modifications Using a Biological Nanopore as a Sensor of Molecular Shape
Orbit 16 (a predecessor model of the Orbit 16 TC instrument) publication in the Journal of the American Chemical Society (2022)
Ensslen T., Sarthak K., Aksimentiev A., Behrends J.C.
2022 - In Vitro Biophysical Characterization of Candidalysin: A Fungal Peptide Toxin
Orbit 16 publication in Frontiers in Methods in Molecular Biology (2022), vol 2542
Lee S., Kichik N., Hepworth O., Richardson J., Naglik J.
2022 - Highly shape- and size-tunable membrane nanopores made with DNA
Orbit 16
Orbit mini and
Vesicle Prep Pro publication in Nature Nanotechnology (2022)
Xing Y., Dorey A., Jayasinghe L., Howorka S.
2022 - Candidalysins Are a New Family of Cytolytic Fungal Peptide Toxins
Orbit 16 publication in mBio (2022)
Richardson J. P., Brown R., Kichik N., Lee S., Priest E., Mogavero S., Maufrais C., Wickramasinghe D. N., Tsavou A., Kotowicz N. K., Hepworth O. W., Gallego-Cortés A., Ponde N. O., Ho J., Moyes D. L., Wilson D., D’Enfert C., Hube B., Naglik J. R.
2022 - A Two-Step Calibration Method for Evaluation High Bandwidth Electrochemical Instrument
Orbit 16 publication in Journal of Electroanalytical Chemistry (2022)
Zhang L.L., Zhong C.B., Li J.G., Niu H.Y., Ying Y.L., Long Y.T.
2022 - A reversibly gated protein-transporting membrane channel made of DNA
Orbit Mini Publication in Nature Communications (2022)
Dey S., Dorey A., Abraham L., Xing Y., Zhang I., Howorka S., Yan H.
2021 - Resolving isomeric posttranslational modifications using a nanopore
Orbit 16 pre-print publication in bioRxiv (2021)
Ensslen T., Sarthak K., Aksimentiev A., Behrends J.C.,
2021 - Design, assembly, and characterization of membrane-spanning DNA nanopores
Orbit 16 and
Orbit mini publication in Nature Protocols (2021)
Lanphere C., Offenbartl-Stiegert D., Dorey A., Pugh G., Georgiou E., Xing Y., Burns J.R., Howorka S.
2020 - Electrophysiology on Channel-Forming Proteins in Artificial Lipid Bilayers
Orbit 16 and
Orbit mini Chapter in Patch Clamp Electrophysiology (2020)
Zaitseva E., Obergrussberger A., Weichbrodt C., Boukhet M., Bernhard F., Hein C., Baaken G., Fertig N., Behrends J.C.
2020 - Electrical recognition of the twenty proteinogenic amino acids using an aerolysin nanopore
Orbit 16 publication in Nature Biotechnology (2020)
Ouldali H., Sarthak K., Ensslen T., Piguet F., Manivet P., Pelta J., Behrends J.C., Aksimentiev A., Oukhaled A.
2020 - Dynamic interaction of fluoroquinolones with magnesium ions monitored using bacterial outer membrane nanopores
Orbit 16 publication in Chemical Science (2020)
Wang J., Prajapati J.D., Kleinekatöfer U., Winterhalter M.
2019 - Synthetic protein-conductive membrane nanopores built with DNA
Orbit 16 and
Orbit mini publication in Nature Communications (2019)
Diederichs T, Pugh G., Dorey A., Xing, Y., Burns J.R., Nguyen Q.H., Tornow M., Tampé R., & Howorka S
2019 - Real-time monitoring β-lactam/β-lactamase inhibitor (BL/BLI) mixture towards the bacteria porin pathway at single molecule level
Orbit 16 publication in Analytical and Bioanalytical Chemistry (2019)
Wang J., Fertig N., Ying Y.L.
2019 - Functional Reconstitution of Membrane Proteins Derived From Eukaryotic Cell-Free Systems
SURFE2R N1 and
Orbit 16 publication in Frontiers in Pharmacology (2019)
Dondapati S.K., Lübberding H., Zemella A., Thoring L., Wüstenhagen D.A., Kubick S.
2019 - Activity of the Gramicidin A Ion Channel in a Lipid Membrane with Switchable Physical Properties
Orbit 16 publication in Langmuir (2019)
Reiter R., Zaitseva E., Baaken G., Halimeh I., Behrends J.C., Zumbuehl A
2019 - A comparison of ion channel current blockades caused by individual poly(ethylene glycol) molecules and polyoxometalate nanoclusters
Orbit 16 publication in The European Physical Journal E (2019)
Wang H., Kasianowicz J.J., Robertson J.W.F., Poster D.L., Ettedgui J.
2018 - The Multifaceted Antibacterial Mechanisms of the Pioneering Peptide Antibiotics Tyrocidine and Gramicidin S
Orbit 16 publication in American Society for Microbiology (2018)
Wenzel M., Rautenbach M., Vosloo J.A., Siersma T., Aisenbrey C.H.M., Zaitseva E., Laubscher W.E., van Rensburg W., Behrends J.C., Bechinger B., Hamoen L.W.
2018 - Size-dependent interaction of a 3-arm star poly(ethylene glycol) with two biological nanopores
Orbit 16 publication in The European Physical Journal E (2018)
Talarimoghari M., Baaken G., Hanselmann R., Behrends J.C.
2018 - Getting drugs into Gram-negative bacteria: Rational rules for permeation through general porins
Orbit 16 publication in ACS Infectious Diseases (2018)
Acosta-Gutierrez S., Ferrara L., Pathania M., Masi M., Wang J., Bodrenko I., Zahn M., Winterhalter M., Stavenger R.A., Pages J.-M., Naismith J.H., van den Berg B., Page M., Ceccarelli M.
2018 - Defined Bilayer Interactions of DNA Nanopores Revealed with a Nuclease-Based Nanoprobe Strategy
Orbit 16 publication in ACS Nano (2018)
Burns J.R., Howorka S.
2018 - Cell‐free production of pore forming toxins: Functional analysis of thermostable direct hemolysin from Vibrio parahaemolyticus
Orbit 16 publication in Engineering in Life Sciences (2018)
Dondapati S.K., Wüstenhagen D.A., Strauch E., Kubick S.
2017 - Validation of ADAM10 metalloprotease as a Bacillus thuringiensis Cry3Aa toxin functional receptor in Colorado potato beetle (Leptinotarsa decemlineata)
Orbit 16 publication in Insect Molecular Biology (2017)
Ruiz-Arroyo V.M., García-Robles I., Ochoa-Campuzano C., Goig G.A., Zaitseva E., Baaken G., Martínez-Ramírez A.C., Rausell C., Real M.D.
2017 - Stability and dynamics of membrane-spanning DNA nanopores
Orbit 16 publication in Nature Communications (2017)
Maingi V., Burns J.R., Uusitalo J.J., Howorka S., Marrink S.J., Sansom M.S.P.
2017 - High-yield production of “difficult-to-express” proteins in a continuous exchange cell-free system based on CHO cell lysates
Orbit 16 publication in Scientific Reports (2017)
Thoring L., Dondapati S.K., Stech M., Wüstenhagen D.A., Kubick S.
2016 - Probing driving forces in aerolysin and α-hemolysin biological nanopores: electrophoresis versus electroosmosis
Orbit 16 publication in Nanoscale (2016)
Boukhet M., Piguet F., Ouldali H., Pastoriza-Gallego M., Pelta J., Oukhaled A.
2016 - Alamethicin Supramolecular Organization in Lipid Membranes from 19F Solid-State NMR
Orbit 16 publication in Biophysical Journal (2016)
Salnikov E.S., Raya J., De Zotti M., Zaitseva E., Peggion C., Ballano G., Toniolo C., Raap J., Bechinger B.
2016 - A biomimetic DNA-made channel for the ligand-controlled and selective transport of small-molecule cargo through a biological membrane
Orbit 16 publication in Nature Nanotechnology (2016)
Burns J.R., Seifert A., Fertig N, Howorka S.
2015 - High-Resolution Size-Discrimination of Single Nonionic Synthetic Polymers with a Highly Charged Biological Nanopore
Orbit 16 and
Vesicle Prep Pro publication in American Chemical Society Nano (2015)
Baaken G., Halimeh I., Bacri, Pelta J., Oukhaled A., Behrends J.C.
2015 - Bilayer-Spanning DNA Nanopores with Voltage- Switching between Open and Closed State
Orbit 16 and
Vesicle Prep Pro publication in American Chemical Society Nano (2015)
Seifert A., Göpfrich K., Burns J.R., Fertig N., Keyser U.F., Howorka S.
2015 - Automated Formation of Lipid Membrane Microarrays for Ionic Single-Molecule Sensing with Protein Nanopores
Orbit 16 publication in Small (2015)
Del Rio Martinez J.M., Zaitseva E., Petersen S., Baaken G., Behrends J.C.
2015 - Antibiotic translocation through porins studied in planar lipid bilayers using parallel platforms
Orbit 16,
Port-a-Patch and
Vesicle Prep Pro publication in Analyst (2015)
Weichbrodt C., Bajaj H., Baaken G., Wang J., Guinot S., Kreir M, Behrends J.C., Winterhalter M., Fertig N.
2014 - Structural and mechanistic insights into the bacterial amyloid secretion channel CsgG
Orbit 16 publication in Nature (2014)
Goyal P., Krasteva P.V., Van GervenN., GubelliniF., Van den BroeckI., Troupiotis-TsaïlakiA., Jonckheere W., Péhau-ArnaudetG., Pinkner J.S., ChapmanM.R., Hultgren S.J., Howorka S., FronzesR., Remaut H.
2014 - Generation of chip based microelectrochemical cell arrays for long-term and high-resolution recording of ionic currents through ion channel proteins
Orbit 16 publication in Sensors and Actuators B: Chemical (2014)
Zheng T., Baaken G., Vellinger M., Behrends J.C., Rühe J.
2013 - Self-Assembled DNA Nanopores That Span Lipid Bilayers
Orbit 16 publication in Nano Letters (2013)
Burns J.R., Stulz E., Howorka S.
2012 - Synthetic Lipid Membrane Channels Formed by Designed DNA Nanostructures
Orbit 16 publication in Science (2012)
Langecker M., ArnautV., Martin T.G., ListJ., RennerS., Mayer M., Dietz H., Simmel F.C.
2011 - Nanopore-based single-molecule mass spectrometry on a lipid membrane microarray
Orbit 16 publication in Journal of the American Chemical Society Nano (2011)
Baaken G., Ankri N., Schuler A.K., Rühe J., Behrends C.
2008 - Planar microelectrode-cavity array for high-resolution and parallel electrical recording of membrane ionic currents
Orbit 16 publication in Lab on a Chip (2008)
Baaken G., Sondermann M., Schlemmer C., Rühe J., Behrends J.C.ポスター
2016 - Parallel and automated formation of lipid bilayers on microstructured chips for ion channel and nanopore recordings
Orbit 16 poster
(1.4 MB)
Kindly provided by Ionera Technologies