Orbit 16 - Taking the Pain out of Painting

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.”

Dr. Friedrich Simmel, Professor, Systems Biophysics and Bionanotechnology, Physics Department and ZNN/WSI
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.“

Dr. Stefan Howorka, Associate Professor of Organic Chemistry and Chemical Biology
University College London, London, UK

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.

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)

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.

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)

Alamethicin - Parallel recordings

  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.

Alpha-Hemolysin - Automated Formation of Membranes from Polyoxazoline based Triblock Copolymers

   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. 

KcsA - Single Channel Recordings

   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.

Ryanodine Receptor - Application of Na-ATP and Ryanodine

   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

MspA - Mycobacterial Porin

   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

Alpha-Hemolysin - PEG Detection

   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

Gramicidin - Ion Channel Forming Antibiotic

   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.

28.06.2018 | Webinar: Artificial Lipid Bilayers in focus: Hand-held DNA-sequencing and biosensing with nanopores

    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 

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.

2018 - Simplifying artificial bilayer experiments: Single-molecule experiments on micro-cavity arrays

   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

2018 - Hand-held DNA-sequencing and biosensing with nanopores

   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

Alpha hemolysin - Lipid bilayer recordings of Alpha hemolysin using the Orbit 16 using the Micro Electrode Cavity Arrays (MECAs) chips

  Orbit 16 application note     (0.4 MB)

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Orbit 16 - Product Sheet

  Orbit 16 product sheet:      (1.4 MB)

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2020 - Electrical recognition of the twenty proteinogenic amino acids using an aerolysin nanopore

  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.

2020 - Dynamic Interaction of Norfloxacin to Magnesium Monitored by Bacterial Outer Membrane Nanopores

  Orbit 16 publication in Pre-Publication in ChemRxiv (2020)

Authors:

Jiajun Wang J., Prajapati J.D., Ulrich Kleinekathöfer U., Winterhalter M.

2019 - Synthetic protein-conductive membrane nanopores built with DNA

   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

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)

Authors:
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)

Authors:
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)

Authors:
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)

Authors:
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)

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.

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)

Authors:
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)

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.

2018 - Defined Bilayer Interactions of DNA Nanopores Revealed with a Nuclease-Based Nanoprobe Strategy

   Orbit 16 publication in ACS Nano (2018)

Authors:
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)

Authors:
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)

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.

2017 - Stability and dynamics of membrane-spanning DNA nanopores

   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.

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)

Authors:
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)

Authors:
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)

Authors: 
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)

Authors: 
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)

Authors: 
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)

Authors: 
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)

Authors: 
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)

Authors:
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)

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.

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)

Authors:
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)

Authors: 
Burns J.R., Stulz E., Howorka S.

2012 - Synthetic Lipid Membrane Channels Formed by Designed DNA Nanostructures

   Orbit 16 publication in Science (2012)

Authors: 
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)

Authors: 
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)

Authors:
Baaken G., Sondermann M., Schlemmer C., Rühe J., Behrends J.C.

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   

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