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ナニオンの Orbit 16 TC(オービット16) のプラットフォームは、一度に16 chの脂質二分子膜を迅速に自動形成し、完全同時測定を行うことで、脂質二分子膜実験における painting法の苦痛から研究者を解放します。 Orbit 16 TC は以下の優位性を有しています:
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 コンプリートシステムは、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。
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.
“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.”
Dr. Friedrich Simmel, Professor, Systems Biophysics and Bionanotechnology, Physics Department and ZNN/WSI
Technical University of Munich, Munich, Germany
“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.“
Dr. Stefan Howorka, Associate Professor of Organic Chemistry and Chemical Biology
University College London, London, UK
Orbit 16 data and applications:
Data courtesy of Dr. Gerhard Baaken et. al., University of Freiburg / Ionera.
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).
Read the full paper. (Am. Chem. Soc Nano, 5(10), 8080-8088, 2011)
Orbit 16 data and applications:
Data courtesy of Dr. Gerhard Baaken et.al, University of Freiburg / Ionera.
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).
Read the full paper: (Am. Chem. Soc Nano, 5(10), 8080-8088, 2011)
Orbit 16 data and applications:
Data courtesy of Dr. Gerhard Baaken, University of Freiburg / Ionera.
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.
Orbit 16 and applications:
Data were kindly provided by Ionera.
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.
(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.
Orbit 16 and applications:
Data were kindly provided by Ionera.
Single channel currents of tetrameric potassium channel KcsA E71A recorded from 5 selected bilayers in parallel.
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.
Orbit 16 and applications:
Data were kindly provided by Ionera.
Traces illustrating RYR single channel activity in the planar lipid bilayer recorded on the Orbit 16.
The RYR channel was reconstituted via fusion of sarcoplasmic reticulum vesicles with preformed asolectin bilayer.
(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.
Conditions: Trans-side: 53 mM Ba(OH)2, 1 mM Ca2+; Cis-side: 150 nM Ca2+, VHold: 0 mV in all cases
Orbit 16 and applications:
Data were kindly provided by Ionera.
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.
Conditions: 20 mM HEPES, 350 mM KCl, pH 7,5, rMspA final concentration 20 ng/ml; holding potential +40 mV
Orbit 16 and applications:
Data were kindly provided by Ionera.
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.
Conditons: 3 M KCl, 20 mM TRIS, pH 8, +40 mV
Orbit 16 and applications:
Data were kindly provided by Ionera.
Screenshots of a recording window of a typical Gramicidin ion channel forming activity assay on the Orbit 16.
Conditions: symmetrical 0,1 HCl, +150 mV.
Orbit mini Webinar
Date: September 17. 2020, 4:00 PM CET (10:00 AM EDT)
Speakers:
Dr. Conrad Weichbrodt (Senior Scientist / Product Manager Orbit family; Nanion Technologies)
Orbit mini and Orbit 16
Learn about single channel measurements in bilayer recording using the orbit instrument family
- Simplifying artificial bilayer experiments: Single-molecule experiments on micro-cavity arrays
- Hand-held DNA-sequencing and biosensing with nanopores
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.
Orbit 16 and Orbit mini Oral Presentation
Presenter:
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
Orbit 16 Oral Presentation
Presenter:
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
Orbit 16 application note (PDF 0.4 MB)
Orbit 16 and Orbit mini publication in Nature Protocols (2021)
Authors:
Lanphere C., Offenbartl-Stiegert D., Dorey A., Pugh G., Georgiou E., Xing Y., Burns J.R., Howorka S.
Orbit 16 and Orbit mini Chapter in Patch Clamp Electrophysiology (2020)
Authors:
Zaitseva E., Obergrussberger A., Weichbrodt C., Boukhet M., Bernhard F., Hein C., Baaken G., Fertig N., Behrends J.C.
Orbit 16 publication in Nature Biotechnology (2020)
Authors:
Ouldali H., Sarthak K., Ensslen T., Piguet F., Manivet P., Pelta J., Behrends J.C., Aksimentiev A., Oukhaled A.
Orbit 16 publication in Chemical Science (2020)
Authors:
Wang J., Prajapati J.D., Kleinekatöfer U., Winterhalter M.
Orbit 16 and Orbit mini publication in Nature Communications (2019)
Authors:
Diederichs T, Pugh G., Dorey A., Xing, Y., Burns J.R., Nguyen Q.H., Tornow M., Tampé R., & Howorka S
Orbit 16 publication in Analytical and Bioanalytical Chemistry (2019)
Authors:
Wang J., Fertig N., Ying Y.L.
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.
Orbit 16 publication in Langmuir (2019)
Authors:
Reiter R., Zaitseva E., Baaken G., Halimeh I., Behrends J.C., Zumbuehl A
Orbit 16 publication in The European Physical Journal E (2019)
Authors:
Wang H., Kasianowicz J.J., Robertson J.W.F., Poster D.L., Ettedgui J.
Orbit 16 publication in American Society for Microbiology (2018)
Authors:
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.
Orbit 16 publication in The European Physical Journal E (2018)
Authors:
Talarimoghari M., Baaken G., Hanselmann R., Behrends J.C.
Orbit 16 publication in ACS Infectious Diseases (2018)
Authors:
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.
Orbit 16 publication in ACS Nano (2018)
Authors:
Burns J.R., Howorka S.
Orbit 16 publication in Engineering in Life Sciences (2018)
Authors:
Dondapati S.K., Wüstenhagen D.A., Strauch E., Kubick S.
Orbit 16 publication in Insect Molecular Biology (2017)
Authors:
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.
Orbit 16 publication in Nature Communications (2017)
Authors:
Maingi V., Burns J.R., Uusitalo J.J., Howorka S., Marrink S.J., Sansom M.S.P.
Orbit 16 publication in Scientific Reports (2017)
Authors:
Thoring L., Dondapati S.K., Stech M., Wüstenhagen D.A., Kubick S.
Orbit 16 publication in Nanoscale (2016)
Authors:
Boukhet M., Piguet F., Ouldali H., Pastoriza-Gallego M., Pelta J., Oukhaled A.
Orbit 16 publication in Biophysical Journal (2016)
Authors:
Salnikov E.S., Raya J., De Zotti M., Zaitseva E., Peggion C., Ballano G., Toniolo C., Raap J., Bechinger B.
Orbit 16 publication in Nature Nanotechnology (2016)
Authors:
Burns J.R., Seifert A., Fertig N, Howorka S.
Orbit 16 and Vesicle Prep Pro publication in American Chemical Society Nano (2015)
Authors:
Baaken G., Halimeh I., Bacri, Pelta J., Oukhaled A., Behrends J.C.
Orbit 16 and Vesicle Prep Pro publication in American Chemical Society Nano (2015)
Authors:
Seifert A., Göpfrich K., Burns J.R., Fertig N., Keyser U.F., Howorka S.
Orbit 16 publication in Small (2015)
Authors:
Del Rio Martinez J.M., Zaitseva E., Petersen S., Baaken G., Behrends J.C.
Orbit 16, Port-a-Patch and Vesicle Prep Pro publication in Analyst (2015)
Authors:
Weichbrodt C., Bajaj H., Baaken G., Wang J., Guinot S., Kreir M, Behrends J.C., Winterhalter M., Fertig N.
Orbit 16 publication in Nature (2014)
Authors:
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.
Orbit 16 publication in Sensors and Actuators B: Chemical (2014)
Authors:
Zheng T., Baaken G., Vellinger M., Behrends J.C., Rühe J.
Orbit 16 publication in Nano Letters (2013)
Authors:
Burns J.R., Stulz E., Howorka S.
Orbit 16 publication in Science (2012)
Authors:
Langecker M., ArnautV., Martin T.G., ListJ., RennerS., Mayer M., Dietz H., Simmel F.C.
Orbit 16 publication in Journal of the American Chemical Society Nano (2011)
Authors:
Baaken G., Ankri N., Schuler A.K., Rühe J., Behrends C.
Orbit 16 publication in Lab on a Chip (2008)
Authors:
Baaken G., Sondermann M., Schlemmer C., Rühe J., Behrends J.C.
Orbit 16 poster (PDF 1.4 MB)
Kindly provided by Ionera Technologies