• Orbit 16

    fully parallel recording of 16 bilayers

Orbit 16 - taking the pain out of painting

Nanion's Orbit 16 platform takes the pain out of bilayer painting and subsequent recordings as it enables fast and successful data generation thanks to the rapid automated formation of 16 bilayers at once and their subsequent fully parallel recording. The benefits the Orbit 16 offers include:

  • Automated formation of 16 artificial lipid bilayers at the push of a button
  • Fully parallel low noise, high bandwidth recordings
  • Substantial data generation due to 16 simultaneous recordings
  • Low noise 16 channel amplifier or integration of external amplifier(s) possible
  • Dedicated comprehensive recording software included
  • Target molecules can be introduced directly or by fusion of (proteo-)liposomes
  • Ion channels: voltage and ligand gated, temperature sensitive
  • Nanopores, antimicrobial peptides, toxins and many more...
  • Cost-efficient experiments with disposable MECA 16 chips

The Orbit 16 is equipped with a dedicated low-noise 16 channel amplifier (Elements s.r.l.) by default enabling the convenient fully parallel recording of 16 bilayers at highest bandwidths. Furthermore, most commercially available amplifiers are compatible with the Orbit 16, including the 16-channel Triton amplifier from Tecella. For single channel amplifiers like the EPC-10 or Axopatch, the Orbit has a built-in multiplexer switchboard, allowing for recordings from the individual bilayers by automatically switching through the recording sites.

The Orbit 16 bilayer platform

A complete Orbit 16 setup consists of the main recording station, an integrated Elements e16 bilayer amplifier and a recording module supporting Ionera's MECA 16 recording chips.

For detailed information:

Software

Orbit 16 EDR Software

eFOUR Software

The software for the Orbit 16 was developed by our partner Elements SRL (Italy). It is intuitive and easy to learn. Please watch this video on the Orbit mini (contains the same software package as the Orbit 16) to learn about the software:

16.07.2015 | Tutorial on Orbit mini

Icon Orbit Mini   Orbit mini

Short introduction on the Orbit mini (tutorial video, 7 minutes)

Consumables

MECA 16

The MECA 16 recording substrate contains a 4 x 4 array of 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 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 are available depending on the sample.

Orbit 16 Meca Chip


Available chip types
  • "Meca 16 Recording Chips 50 µm": 16-well recording chip with 50 µm cavity size (Order # 131002)
  • "Meca 16 Recording Chips 16 µm": 16-well recording chip with 16 µm cavity size (Order # 131003)

Testimonials & Case Studies

Prof. Dr. Friedrich Simmel - Statement about the Orbit 16

Icon Orbit   “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

Icon Orbit   “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

Data and Applications

Alamethicin - Parallel recordings

Alamethicin IoneraIcon Orbit   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 - PEG Detection

Icon Orbit   Orbit16 Ionera ahemolysinOrbit 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

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

Icon Orbit   Orbit16 Ionera ahemolysin 2Orbit 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. 

Alpha-Hemolysin - Block by Mono- and Poly PEGs

Icon Orbit   2011 HL HistOrbit 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)

Alpha-Hemolysin - Parallel Recordings of monoPEG-28 Block

2011 HemolysinIcon Orbit   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)

Gramicidin - Ion Channel Forming Antibiotic

Icon Orbit   Orbit16 Ionera GramicidinOrbit 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.

KcsA - Single Channel Recordings

Icon Orbit   Orbit16 Ionera KcsAOrbit 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.

MspA - Mycobacterial Porin

Icon Orbit   Orbit16 Ionera MspAOrbit 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

Ryanodine Receptor - Application of Na-ATP and Ryanodine

Icon Orbit   Orbit 16 RyanodineR 1Orbit 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

Webinars and Movies

27.01.2016 | Webinar: Instant bilayers - just add protein.

Icon Orbit   Orbit 16 and   Icon Orbit Mini   Orbit Mini

Orbits V1 flat 250pxThis 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.

 

Downloads:

Application Notes

Product Sheets

Orbit 16 - Product Sheet

Icon Orbit   Orbit 16 product sheet:   logo pdf   (1.4 MB)

Publications

2017 - Validation of ADAM10 metalloprotease as a Bacillus thuringiensis Cry3Aa toxin functional receptor in Colorado potato beetle (Leptinotarsa decemlineata)

Icon Orbit  Orbit 16 publication in Insect Molecular Biology (2017)

2017 - High-yield production of “difficult-to-express” proteins in a continuous exchange cell-free system based on CHO cell lysates

Icon Orbit   Orbit 16 publication in Scientific Reports (2017)

2016 - Probing driving forces in aerolysin and α-hemolysin biological nanopores: electrophoresis versus electroosmosis

Icon Orbit  Orbit 16 publication in Nanoscale (2016)

2016 - Alamethicin Supramolecular Organization in Lipid Membranes from 19F Solid-State NMR

Icon Orbit  Orbit 16 publication in Biophysical Journal (2016)

2016 - A biomimetic DNA-made channel for the ligand-controlled and selective transport of small-molecule cargo through a biological membrane

Icon Orbit  Orbit 16 publication in Nature Nanotechnology (2016)

2015 - High-Resolution Size-Discrimination of Single Nonionic Synthetic Polymers with a Highly Charged Biological Nanopore

Icon Orbit  Orbit 16 and   icon vpp   Vesicle Prep Pro publication in American Chemical Society Nano (2015)

2015 - Bilayer-Spanning DNA Nanopores with Voltage- Switching between Open and Closed State

Icon Orbit  Orbit 16 and   icon vpp   Vesicle Prep Pro publication in American Chemical Society Nano (2015)

2015 - Automated Formation of Lipid Membrane Microarrays for Ionic Single-Molecule Sensing with Protein Nanopores

Icon Orbit  Orbit 16 publication in Small (2015)

2015 - Antibiotic translocation through porins studied in planar lipid bilayers using parallel platforms

Icon Orbit  Orbit 16,   icon pap   Port-a-Patch and   icon vpp   Vesicle Prep Pro publication in Analyst (2015)

2014 - Structural and mechanistic insights into the bacterial amyloid secretion channel CsgG

Icon Orbit  Orbit 16 publication in Nature (2014)

2014 - Generation of chip based microelectrochemical cell arrays for long-term and high-resolution recording of ionic currents through ion channel proteins

Icon Orbit  Orbit 16 publication in Sensors and Actuators B: Chemical (2014)

2013 - Self-Assembled DNA Nanopores That Span Lipid Bilayers

Icon Orbit   Orbit 16 publication in Nano Letters (2013)

2012 - Synthetic Lipid Membrane Channels Formed by Designed DNA Nanostructures

Icon Orbit   Orbit 16 publication in Science (2012)

2011 - Nanopore-based single-molecule mass spectrometry on a lipid membrane microarray

Icon Orbit   Orbit 16 publication in Journal of the American Chemical Society Nano (2011)

2008 - Planar microelectrode-cavity array for high-resolution and parallel electrical recording of membrane ionic currents

Icon Orbit  Orbit 16 publication in Lab on a Chip (2008)

Posters

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Nanion Technologies GmbH

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D-80339 Munich - Germany
info@nanion.de