高通量脂双层工作站

Orbit 16 TC - 高通量脂双层工作站

Nanion公司的Orbit 16 TC 平台可轻松高效的进行脂双层涂抹与记录，该设备能自动一次性形成16个脂双层并对其进行完全平行记录，所以能快速地生成有效的数据。

Orbit 16 TC 主要特点有：

• 一键操作自动形成16个人工脂双层
• 完全平行的低噪音、高带宽记录
• 可同时记录16个脂双层从而获得大量的数据
• 低噪音16通道放大器，也可以兼容外置放大器
• 附带专用记录软件
• 目标蛋白可直接插入和通过蛋白脂质体插入
• 可记录: 电压、配体门控离子通道、温度敏感离子通道
• 也可记录纳米孔、抗菌肽、毒素等...
• 低实验成本的MECA 16 TC芯片

Orbit 16 TC 内置专业的低噪16通道放大器(Elements s.r.l.)，支持使用最高带宽同时记录16个脂双层。此外，Orbit 16 TC 还兼容绝大多数在售的放大器，包括Tecella公司的16通道Triton放大器、EPC-10或Axopatch等单通道放大器等。Orbit内置多路转接器，可以自动在记录位点之间切换从而实现用单通道放大器记录多个脂双层位点。

Orbit 16 TC 脂双层工作站

完整的Orbit 16 TC 系统包括主记录站、内置的Elements e16脂双层专用放大器和支持Ionera公司的MECA 16 TC 16记录芯片的记录模块。

Orbit 16 TC EDR4 Software

MECA 16 TC 耗材
MECA 16 TC记录基板在高惰性聚合物中包含16个圆形微腔阵列。每个腔体包含一个单独的集成Ag/AgCl微电极。双分子层通过远程驱动喷涂（Ionera1扩散）自动形成，这样就可以覆盖充满液体的空腔。双层膜可以以自动化的方式轻松重复地被破坏和重新成形。双层形成后，离子通道或纳米孔通过自插入、蛋白脂质体融合或洗涤剂胶束稀释来重建。

数据与应用：

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 case

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.


发表文献：
2022 - Candidalysins Are a New Family of Cytolytic Fungal Peptide Toxins
2021 - Design, assembly, and characterization of membrane-spanning DNA nanopores
2021 - Resolving isomeric posttranslational modifications using a nanopore
2020 - Electrophysiology on Channel-Forming Proteins in Artificial Lipid Bilayers
2020 - Electrical recognition of the twenty proteinogenic amino acids using an aerolysin nanopore
2020 - Dynamic interaction of fluoroquinolones with magnesium ions monitored using bacterial outer membrane nanopores