Orbit mini - プラグ＆ペイント

Prof. Dr. Chris Miller - Case Study about the Orbit mini

   "Contrary to my initial expectations, the Orbit mini (red arrow) has performed exceptionally well for
our application, with vastly improved signal-tonoise characteristics over our best recordings on the home-built system."

“The compactness of the instrument makes for flexible and convenient placement in the lab. With a tiny footprint, it sits on a table or desk accompanied by only a laptop. In addition, bilayers are painted 'blind' with a teflon wand supplied by Nanion without observing the partition through a microscope. Thus, the
entire support-package of microscope, illuminator, Faraday cage, vibration-table, and rig-rack is eliminated. The acquisition software is intuitive and robust, and the data can be stored in formats readable by the Axon analysis programs that we routinely use. Moreover, the four electrically independent holes in the chip-partition means that four bilayers can be monitored simultaneously, thus quadrupling the probability of catching a clean single channel to record. This represents a significant, practical boost in experimental efficiency.”

Source: "Getting going with the Orbit mini planar bilayer system: A narrative"

From:
Prof. Dr. Chris Miller, Professor of Biochemistry, Investigator, Howard Hughes Medical Institute Member, US National Academy of Sciences, Brandeis University, Waltham, Massachusetts, USA

Read the full customer case study here

Alpha-Hemolysin - Temperature Control

   Orbit mini and applications:
Data were kindly provided by Ionera.

Temperature control of the Orbit mini: PEG induced current blockages of a alpha-hemolysin pore

(A) Current traces recorded at 10°C and 40°C illustrating a strong increase of the open pore current as well as the blockage frequency at elevated temperature.
(B) Event averaged histograms of the residual current during blockages. The open pore current scales with the temperature as well as the dwell time of the blockages.
(C) Dependence of the open pore current on the temperature.
(D) Dependence of the frequency of blockages on the temperature.

OccK1 - Outer Membrane Protein of the Pathogenic Bacterium Pseudomonas Aeruginosa

   Orbit mini and applications:
Data were kindly provided by Ionera.

Representative recording from a single OmpF trimer showing current blocks due to the translocation of antibiotic Enrofloxacin, event averaged histogram.
Conditions: 150 mM KCl, 5 mM MES, pH 6, 10 mM enrofloxacin, + 60 mV

OmpF - Current block

   Orbit mini and applications:
Data were kindly provided by Ionera.

Representative recording from a single OmpF trimer showing current blocks due to the translocation of antibiotic Enrofloxacin, event averaged histogram.
Conditions: 150 mM KCl, 5 mM HEPES, 5 mM MES, pH 6, 10 mM enrofloxacin, +60 mV

TRPA1 - Temperature Dependency

   Orbit mini data and applications:

(A) Effect of temperature on TRPA1 activity
(B) The open probability (Po) versus the temperature and fitted with Boltzmann equation (EC₅₀ was found at 14˚C).

The Arrhenius plot of the same data resulted in a Q10 of 46 (Literature: Q10 ~ 40).

VDAC - Application of Fluoxetine

   Orbit mini and applications:
Data were kindly provided by Ionera.

Current recordings of single VDA channels from selected bilayers in parallel.
VDAC was reconstituted in DPhPC membranes and partially blocked upon the addition of Fluoxetine (Prozac).

(A) Representative parallel recording from 3 VDAC channels in DPhPC membranes. Conditions: 1 M KCl, 25 mM Tris, pH 7,5, voltage change protocol is given below.
(B) Current trace illustration partial blocade of VDAC by Fluoxetine (Prozac).
(C) Current-voltage trace of a single VADC channel under a ramping voltage of -70/+70 mV before (green) and after (blue) addition of 50 µm Fluoxetine

Purified VDAC sample is a generous gift from Prof. Stephan Nussberger to Ionera, Department Biophysics, University of Stuttgart, Germany

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

2015 - Orbit mini Membrane Preparation Tutorial Video

   Orbit Mini

Short introduction how to prepare membranes on the Orbit mini

2015 - Orbit mini Tutorial Video

   Orbit mini

Short introduction on the Orbit mini

17.09.2020 | Webinar: Electrophysiological investigation of integral membrane proteins using the Orbit mini

  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)

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.

TRPV1 - "Temperature activation of TRPV1 reconstituted into planar lipid bilayers and recorded using the Orbit mini platform"

  Orbit mini application note      (0.4 MB)

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Orbit mini - Product Sheet

   Orbit mini product sheet      (1.2 MB)

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Orbit mini Product Flyer - Temperature Control

  Orbit mini Product flyer     (0.4 MB)

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Orbit mini - Quickstart Guide "Getting Started"

   Orbit mini Quickstart Guide "Getting Started"

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2021 - Design, assembly, and characterization of membrane-spanning DNA nanopores

   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.

2021 - ATP and large signaling metabolites flux through caspase-activated Pannexin 1 channels

   Orbit mini publication in eLife (2021)

Authors:
Narahari A.K., Kreutzberger A JB., Gaete P.S., Chiu Yu-Hsin, Leonhardt S.A., Medina C.B., Jin X., Oleniacz P.W., Kiessling V., Barrett P.Q., Ravichandran K.S., Yeager M., Contreras J.E., Tamm L.K., Bayliss D.A.

2020 - Structure and mechanism of bactericidal mammalian perforin-2, an ancient agent of innate immunity

   Orbit mini and      Vesicle Prep Pro publication in Science Advances (2020)

Authors:
Ni T., Jiao F., Yu X., Aden S., Ginger L., Williams S.I., Bai F., Pražák V., Karia D., Stansfeld P., Zhang P., Munson G., Anderluh G.,Scheuring S., Gilbert R.J.C.

2020 - Small Molecule Permeation Across Membrane Channels: Chemical Modification to Quantify Transport Across OmpF

   Orbit mini pre-publication in Angewandte Chemie (2020)

Authors:
Wang J., Bafna J.A., Bhamidimarri S.P., Winterhalter M.

2020 - New Structural insights into Kir channel gating from molecular simulations, HDX-MS and functional studies

  Orbit mini publication in Nature Scientific Reports (2020)

Authors: 
Fagnen C., Bannwarth L, Oubella I., Forest E., De Zorzi R., de Araujo A., Mhoumadi Y., Bendahhou S., Perahia D., Vénien-Bryan C.

2020 - Electrophysiology on Channel-Forming Proteins in Artificial Lipid Bilayers

  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.

2020 - Electrophysiological characterization of transport across outer membrane channels from Gram‐negative bacteria in presence of lipopolysaccharides (LPS)

   Port-a-Patch,     Vesicle Prep Pro and      Orbit mini Publication in Angewandte Chemie International Edition (2020)

Authors:
Wang J., Terrasse R., Bafna J.A., Benier L., Winterhalter M.

2020 - Cryo-EM structure of human Cx31.3/GJC3 connexin hemichannel

  Orbit mini publication in Science Advances (2020)

Authors:
Lee H.J., Jeong H., Hyun J., Ryu B., Park K., Lim H.H., Yoo J., Woo J.S.

2020 - Computational Modeling of Ion Transport in Bulk and through a Nanopore Using the Drude Polarizable Force Field

   Orbit mini publication in Journal of Chemical Information and Modeling (2020)

Authors:
Prajapati J.D., Mele C., Alphan Aksoyoglu M.A., Winterhalter M., Kleinekathöfer U.

2020 - Aerolysin nanopores decode digital information stored in tailored macromolecular analytes

   Orbit mini publication in Science Advances (2020)

Authors:
Cao C., Krapp L.F., Al Ouahabi A., König N.F., Cirauqui N., Radenovic A., Lutz J.F. Dal Peraro M.

2020 - Activation of the archaeal ion channel MthK is exquisitely regulated by temperature

  Orbit mini publication in eLife (2020)

Authors: 
Jiang Y., Idikuda V., Chowdhury S., Chanda B.

2019 - Structure of the human ClC-1 chloride channel

   Port-a-Patch,     Vesicle Prep Pro and      Orbit mini publication in PLOS Biology (2019)

Authors:
Wang K., Preisler, SS, Zhang, L., Cui, Y., Missel, JW., Grønberg C., Gotfryd, K., Lindahl E., Andersson, M., Calloe, K., Egea P.F., Klaerke D.A., Pusch M., Pedersen P.A., Zhou, Z.H., Gourdon, P.

2019 - Revisiting an old antibiotic: bacitracin neutralizes binary bacterial toxins and protects cells from intoxication (2)

   Orbit mini publication in The FASEB Journal (2019)

Authors:
Schnell L., Felix I., Müller B., Sadi M., von Bank F., Papatheodorou P., Popoff M.R., Aktories K., Waltenberger E., Benz R., Weichbrodt C., Fauler M., Frick M., Barth H.

2019 - Purification of Functional Human TRP Channels Recombinantly Produced in Yeast

   Orbit mini publication in Cells (2019)

Authors:
Zhang L., Wang K., Klaerke D.A., Calloe K., Lowrey L., Pedersen P.A., Gourdon P., Gotfryd K.

2019 - Arg-8 of yeast subunit e contributes to the stability of F-ATP synthase dimersand to the generation of the full-conductance megachannel

    Orbit mini publication in Journal of Biological Chemistry (2019)

Authors:
Guo L., Carraro M., Carrer A., Minervini G., Urbani A., Masgras I., Tosatto S.C.E., Szabò I., Bernardi P., Lippe G.

2018 - Structures of monomeric and oligomeric forms of the Toxoplasma gondii perforin-like protein 1

   Orbit mini and      Vesicle Prep Pro publication in Science Advances (2018)

Authors:
Ni T., Williams S.I., Rezelj S., Anderluh G., Harlos K., Stansfeld P.J., Gilbert R.J.C.

2018 - Functional Analysis of Membrane Proteins Produced by Cell-Free Translation

   Port-a-Patch and      Orbit mini article in Protein Engineering (2018)

Authors:
Dondapati S.K., Wüstenhagen D.A., Kubick S.

2018 - Dynamics and number of trans-SNARE complexes determine nascent fusion pore properties

   Orbit mini publication in Nature (2018)

Authors:
Bao H., Das D., Courtney N.A., Jiang Y., Briguglio J.S., Lou X., Roston D., Cui Q., Chanda B., Chapman E.R.

2017 - Molecular determinants of permeation in a fluoride-specific ion channel

   Orbit mini in eLife (2017)

Authors:
Last .B., Sun S., Pham M.C., Miller C.

2017 - Engineering a pH responsive pore forming protein

  Orbit mini and      Vesicle Prep Pro publication in Scientific Reports (2017)

Authors: 
Kisovec M., Rezelj S., Knap P., Cajnko M.M., Caserman S., Flašker A., Žnidaršič N., Repič M., Mavri J., Ruan Y., Scheuring S., Podobnik M., Anderluh G.

2016 - Mechanistic signs of double-barreled structure in a fluoride ion channel

  Orbit mini publication in eLIFE (2016)

Authors:
Last N.B., Kolmakova-Partensky L., Shane T., Miller C.

2016 - Next level toxicity screening: From single channel to overall cell behavior

  Orbit mini,      CardioExcyte 96 and      SyncroPatch 384PE (a predecessor model of the SyncroPatch 384i) poster, Meeting of the French Society of Toxinology (SFET) 2015     (0.9 MB)

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