パッチクランプのすべてのニーズに対応

SyncroPatch 384i システム



最先端の自動分注機Biomek i5にパッチクランプモジュールを統合した、画期的な自動パッチクランプシステムです。384細胞同時測定を行い、1日あたり20,000データポイントのスループットを実現します。その使いやすさとオープンなデザインで、完全な自動化とHTSへの統合をサポートします。

また32ウェルモードを使用することにより、ハイスループットスクリーニングに限らず、小規模なスクリーニングプロジェクトや学術研究など、どのようなスループットのニーズにも対応することができ、NPC-384チップのコストメリットを最大限に活用することができます。

実験に必要なウェル数を32の倍数で選択し、残りのウェルを数日間にわたって使用することができます。またプログラムを完全に自動化することにより、8時間以上の実験を無人で行うこともできます。

卓越したデータ品質と柔軟性により、SyncroPatch 384は、グローバルな製薬会社、CRO、学術機関などで採用されています。


主な特徴 

  • 384細胞同時測定
  • 32ウェルモードにより小規模な化合物スクリーニングや研究プロジェクトにも最適化
  • 高いギガシール成功率
  • 高速外部溶液交換(最大110 µl/s)が可能
  • 記録中の内部溶液の灌流が可能(例:細胞内カルシウム依存性のK+チャネルの記録が可能)
  • 測定部と12個のデッキを個別に温度コントロール可能(温度範囲:10~37℃)
  • カレントクランプによる活動電位の測定も可能
  • シングル & マルチホールチップを使用可能(自社製造)
  • プログラムの自動化により、約8時間の無人運転が可能
  • 濃度解析のために測定後の溶液を回収することが可能
  • 優れたサービスとサポートを提供します

ハードウェアー/ソフトウェア

  • ナニオンのパッチクランプチップは、マニュアルパッチクランプで使用されるガラスキャピラリーと同様にホウケイ酸ガラス製のため、低キャパシタンス、低化合物吸着、優れたパッチ/シール特性を実現しています
  • オンデッキでの化合物プレートの準備 - 化合物の準備のために別の自動分注機を使用したり、手作業で化合物プレートを準備する必要はありません。
  • 流路タイプのチップと比較して、溶液と壁面の接地面積が低いため、化合物の吸着が低い
  • 化合物暴露 & wash回数の制限なし
  • グリッパーとピペッティングヘッドが独立して稼働するため、例えば化合物の蓋を外す際にピペッティングチップを外す必要がありません
  • 少量の化合物で実験が可能
  • 強力な解析ソフトウェア(フレキシブルなバッチ解析、IC50計算、IV解析、Genedata Screenerとの互換性あり)
  • z-primeとヒートマップにより、データの概要を迅速かつ視覚的に把握することが可能
  • 設定、編集、分析が簡単にできます
  • スタンダードとアドバンスの異なる操作モードにより、実験パラメータの柔軟性を維持しながら、シンプルに操作できます
  • スループットの選択 - 32ウェルモードでは、実験に適したスループットを常に選択できます

アプリケーション

  • 電位依存性 & リガンド依存性チャネルに対応
  • 少ない細胞量で実験が可能 – 幹細胞や初代細胞にも適用可能
  • 様々なイオンチャネルに適用可能
  • CiPA validation studyにおいて様々なデータを提供
  • 創薬のあらゆるフェーズに対応
  • データを様々なフォーマットでエクスポートし、外部ツールで簡単に解析可能
  • 様々なCROアッセイに対応できる柔軟性



特徴と仕様

Technical Specifications of the SyncroPatch 384
SyncroPatch 384: Standard Delivery Package

The SyncroPatch 384 includes:

  • Biomek i5 with a 384-pipettor arm, gripper and temperature-controlled deck positions
  • Temperature-controlled patch clamp module
  • Amplifier (384 channels) incl. current clamp
  • Windows 10 OS with PatchControl 384 and DataControl 384 software suite
  • Guided Labware Setup and Method Launcher
  • Temperature-controlled cell hotel
  • Barcode scanner
  • NPC-384 borosilicate recording chips
  • Optional service plans for unmatched



ソフトウェア

PatchControl 384
PatchControl 384 is a powerful graphical user interface for intuitive, quick and easy setup of voltage protocols and experimental parameters. The recording wells are visualized and color-coded based on user-defined quality criteria, e.g. seal resistance, series resistance or capacitance. With one mouse click, the view switches to online analysis results, for example I/V curves or concentration-response curves.
DataControl 384: The Analysis Software
DataControl 384 is used to visualize and analyze the PatchControl 384-data, employing user-defined data analysis templates. Results (automated IC50, EC50, IV relationship plot generation), compound information, and quality control parameters are exported together in a user-defined export format, automatically generating pdf-reports, and preparing the data for further database integration. This process is straightforward, intuitive and quickly accomplished.



専用チップ

NPC-384

The NPC-384 chip is the cost-efficient and high quality consumable for the SyncroPatch 384. It is produced and quality-assured in-house at Nanion headquarters in Munich. Different types of NPC-384 chips are available which should be chosen depending on cell size and application.

Material

The borosilicate glass slide with the patch aperture is encased in a 384 well plate forming wells where the cells and external solutions are delivered. The design of the chip allows perfusion of the internal solution during an experiment.

Features

Each NPC-384 chip contains 384 recording chambers. These sites can be used all at one time or in 32-well mode, parts of the chip can be used in multiples of 32, and the remaining parts used over several days, without a reduction success rate. One chip can be measured on the SyncroPatch 384 and 25 chips can be stacked in the robot for unattended experiments. The open design of the chip makes sample collection and subsequent verification of compound concentration. Additionally, the number of exchanges of either the internal or the external solution is unlimited. NPC-384 chips can be purchased as single hole per well for GOhm seals, or multi-hole per well to increase measured current amplitude and improve success rate.

Available chip types
  • "NPC-384, 1x medium resistance": One hole per well (Order # 221102)
  • "NPC-384, 1x medium resistance plus": One hole per well (Order # 221104)
  • "NPC-384, 4x medium resistance": 4 holes per well (Order # 221402)
  • "NPC-384, 1x high resistance": One hole per well (Order # 221101)
  • "NPC-384, 4x high resistance": 4 holes per well (Order # 221401)
  • "NPC-384, 1x low resistance": One hole per well (Order # 221103)
  • "NPC-384, 4x low resistance": 4 holes per well (Order # 221403)
  • "NPC-384, 8x": 8 holes per well (Order # 221801)



試薬

Buffers and Solutions for the SyncroPatch 384/384i/768i

Reliable buffer solutions are critical for any electrophysiological application. Our goal, therefore, is to provide high-class recording solutions that leave you in no doubt about quality and stability. Our quality assurance includes chemical tests as well as tests on our patch clamp systems of each lot. Our buffers are shipped with the corresponding "Certificates of Analysis" and "Material Safety Data Sheets" (MSDS).

Available buffers and solutions
  • "External Standard", 500 mL: (Order # 08 3001)
  • "External Standard Ca 10", 500 mL: (Order # 08 3012)
  • "External NMDG 60", 500 mL: (Order # 08 3004)
  • "External NMDG 60 Ca 10", 500 mL: (Order # 08 3011)
  • "External [-] Ca2+ [-] Mg2+", 500 mL: (Order # 08 3003)
  • "Internal CsF 110", 500 mL: (Order # 08 3008)
  • "Internal KF 110", 500 mL: (Order # 08 3007)
  • "Washing solution", 5 L: (Order # 08 3010)



データ&アプリケーション

32- well mode for smaller screens or academic investigations

SyncroPatch 384 data and applications:
Cells were kindly provided by SB Drug Discovery

An exemplary 32-well Mode Experiment. A small fraction of the chip can be used at a time, which is ideal for smaller compound screens.
Consecutive experiments of 32-wells on the same NPC-384 patch clamp chip over multiple days. Success rate and accurate pharmacology remains stable over 8 days as shown in the figure. Nav1.5 recordings in the presence of increasing Mexiletine concentrations.


AMPA Receptor (GluA2) - Activation by Glutamate

SyncroPatch 384PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by SB Drug Discovery.

The AMPA receptor (GluA2) was activated using different concentrations of glutamate (1 µM - 100 µM). Measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384), the whole cell patch methodology and multi-hole chips were used.
The lower two images are displaying screenshots of single cell currents after repetitive glutamate applications:
Left: The same concentration of Glutamate was applied three times.
Right: Four different Glutamate concentrations were applied in a cumulative manner.


AMPA Receptor (GluA2) - Cumulative Concentration Response

SyncroPatch 384PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by SB Drug Discovery.

The AMPA receptor (GluA2)was activated by increasing concentrations of glutamate on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384). L-glutamate was applied for approximately 500 ms in increasing concentrations (A) and a cumulative concentration response curve for glutamate was constructed for 222 wells (C).
The online analysis values peak amplitude and area under the curve (AUC) are shown versus time in Panel B. The fast activation of GluA2 could be captured at higher concentrations (inset; 1 mM).


AMPA Receptor (GluA2) - Pharmacology

SyncroPatch 384PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by SB Drug Discovery.

The AMPA receptor (GluA2) was analyzed using different positive and negative allosteric modulators (CNQX, LY404187, LY395153, CP465022, Cyclothiazide). After activating the receptor by application of Glutamate, the modulating compound plus glutamate was applied afterwards. Measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384), the whole cell patch methodology and multi-hole chips were used.
The lower images on the left hand side are displaying a screenshot of a current after application of the positive modulator LY404187. The EC50 was determined as 379 nM.


Cardiac Ion Channels - Pharmacology of Sotalol

CardioExcyte 96 and SyncroPatch 384PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River and Cellular Dynamics.

The image on the left hand side displays the results of the blocking effect of Sotalol on hERG. The result is in good agreement with manual patch clamp data (Crumb et al., 2016). The compound induced arrhythmia when iPSC-CM were exposed to a minimum concentration of 10 µM. Arrhytmic events were both detected in field potential recordings as well as in the impedance based contractility measurements.


CaV1.2 - Current Voltage Relationship

SyncroPatch 384PE (a predecessor model of SyncroPatch 384) data and applications:
Cells kindly provided by Charles River.

CaV1.2 expressed in CHO cells recorded on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384). A The screenshot shows the data acquisition and analysis software used on the SyncroPatch 384PE. The online analysis values are shown for a current-voltage experiment. B The raw traces from an example cell elicited by depolarizing steps from -60 mV to 40 mV in 10 mV increments from a holding potential of -80 mV are shown. C The normalized current-voltage plot for an average of 272 cells. A Boltzmann equation fit revealed a V0.5 of activation of -4.8 mV.


CaV1.2 - Pharmacology of Nifedipine, using the CiPA protocol

SyncroPatch 384PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.

Screenshots of the PatchControl 384 software showing hCaV1.2/β2/α2δ1 current traces in response to the CiPA voltage step protocol and the corresponing current-voltage relationship plot. Measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384) using perforated patch methodology (Escin) and multi-hole chips (4 holes per well), the success rate of valuable data for the analysis was 94%. The IC50 value of Nifedipine was determined as 106 nM.


CaV1.2 - Stable recording from frozen stock cells

SyncroPatch 384PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.
Screenshots of the PatchControl 384 software showing hCaV1.2β2/α2δ1 current traces in response to a voltage step protocol and the corresponding current-voltage relationship plot. Measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384) using perforated patch methodology (Escin) and multi-hole chips (4 holes per well), the success rate of valuable data for the analysis was 100 %. The cells were used from a frozen cell stock (after induction) and recorded stably for more than 20 minutes. The IC50 value of Nifedipine was determined as 21 nM.

ClC-1 - Current-voltage plot

SyncroPatch 384i (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.

Activation of hClC-1 tail currents expressed in CHO cells recorded on the SyncroPatch 384i (a predecessor model of SyncroPatch 384). A pre-pulse voltage step to +60 mV was followed by voltage steps from -120 mV to +80 mV for 300 ms (increasing in 20 mV steps) and the tail current was measured at the subsequent step to -100 mV. Out of a possible 384 wells, all 384 wells were used for the IV analysis


ClC-1 – Inhibition by 9-AC

SyncroPatch 384i (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.

Tail currents of ClC-1 expressed in CHO cells were inhibited by increasing concentration of 9-AC. A single concentration of 9-AC was added to each well and the concentration response curve constructed over multiple wells. The IC50 was calculated to be 6.3 µM for an average of 352 wells. The average current traces are also shown.


GABAA Receptor (α1β2γ2) - Success Rates

SyncroPatch 384PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Bsys.
Statistic of hGABAA α1β2γ2 cells recorded on one NPC-384 1-hole (1x) patch clamp chip. 57 % of the cells on one NPC-384 chip had seal resistance > 1 GOhm at the beginning and 48% at the end of the experiment. Access (RSeries) was good with 80% of cells with RSeries <20 MOhm at the start of the experiment.

GluA2 activation at 110µl/s – speed is key

SyncroPatch 384 data and applications:
Cells were kindly provided by SB Drug Discovery

The AMPA receptor (GluA2) was activated using increasing concentrations of glutamate. Measured on the SyncroPatch 384 the whole cell patch methodology and multi-hole chips were used. The faster you apply the ligand, the shorter is the Time to Peak, this means pipetting speed is relevant for accurate pharmacology. The IC50 of Glutamate at 110 µl/s was 460 µM.


Glycine Receptor (GlyRa1) - Reproducible Current Recordings

SyncroPatch 384PE (a predecessor model of SyncroPatch 384) data and applications:
Glycine-mediated current traces and corresponding time plots from 384 simultaneously recorded HEK cells are shown. Multiple additions of 50 µM Glycine produce very robust current responses with similar peaks, providing best conditions for cumulative pharmacology on one cell.

hERG - Pharmacology at Physiological Temperature using the CiPA Protocol

SyncroPatch 384/768 PE (a predecessor model of the SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.
Screenshots of the PatchControl 384 software showing hERG current traces in response to the CiPA voltage step protocol at 35 degree Celsius. Measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384) using perforated patch clamp methodology (Escin) and multi-hole chips (4 holes per well). The IC50 value of Erythromycin of the peak current was determined as 60.5 µM. 

hERG - Pharmacology using the CiPA Protocol

SyncroPatch 384/768 PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.
Screenshots of the PatchControl 384 software showing hERG current traces in response to the CiPA voltage step protocol. Measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384) using whole cell patch clamp methodology and multi-hole chips (4 holes per well). The IC50 value of the following compounds of the peak current was determined as 4.18 µM for Diltiazem, 37.4 nM for Terfenadine, 971 nM for Quinidine, 63 µM for Mexiletine, 431 nM for Verapamil and 4.54 µM for Ranolazine. 

hERG - recordings with great stability using the CiPA step ramp protocol

SyncroPatch 384PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.
Screenshots of the PatchControl 384 software showing hERG current traces in response to the CiPA voltage step protocol. Measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384) using perforated patch clamp methodology (Escin) and multi-hole chips (4 holes per well). 

hERG - Stable Recordings with Accurate Pharmacology

SyncroPatch 384PE (a predecessor model of SyncroPatch 384) data and applications: 
Cells were kindly provided by Charles River Laboratories.
Current-voltage relationship of hERG (Kv11.1) expressed in HEK293 is shown along with pharmacology of 4 hERG-active compounds. The current-voltage relationships for all 384 wells (top) using perforated patch (Escin) and multi-hole chips (4 holes per well) are shown. In all 384 wells, a hERG-mediated current was observed with peak amplitude >700 pA at -20 mV. Using a pharmacology voltage protocol, experiments were stable lasting over 20 minutes. Concentration response curves for astemizole, pimozide, cisapride and terfenadine revealed IC50 values consistent with those found in the literature. 

hERG and Temperature Control

SyncroPatch 384 data and applications:
Cells were kindly provided by Charles River Chantest
Cardiac ion channels are recommended to be recorded at Phys. Temp. (ICH S7B Q&A. 2021). On the SyncroPatch 384, measurement site, cells and solutions can be accurately temperature controlled – in the presence of physiological temperatures the hERG current kinetic is changed to a larger slope and higher amplitude.

KCa1.1 (BK) - High throughput study

SyncroPatch 384i (a predecessor model of SyncroPatch 384) data and applications:
Data kindly provided by Sharan R. Srinivasan1 and Vikram G. Shakkottai1,2
1Department of Neurology, University of Michigan, Ann Arbor, MI 48109;
2Molecular and Integrative Physiology, University of Michigan, Ann Arbor, MI 48109.
HEK293 cells stably transfected with BK channels were used to screen over 50,000 compounds, and using clever buffering techniques, targeting only activators of calcium sensitivity for BK channel augmentation. 

KCa3.1 (SK4) - Activation by Perfusion of free internal Calcium

SyncroPatch 384/768 PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.
Screenshots of the PatchControl 384 software are showing KCa3.1 raw traces and according time plots (online analysis) to a voltage ramp from -120 mV to + 60 mV over 200 ms. The application of internal Ca2+ is indicated by the yellow bar. The current increased upon application of internal Ca2+ reaching a peak within 1-2 min after the start of the perfusion. Five minutes of stable KCa3.1 current was recorded prior the channel was inhibited by cumulative additions of external Ba2+; first partly (1 mM Ba2+) and then completely (5 mM Ba2+). The recording was performed with perfectly high success rates in whole cell configuration on a multi hole chip (4 holes per well) using the SyncroPatch 384PE (a predecessor model of SyncroPatch 384).

Kir2.1 - Pharmacology of Barium

SyncroPatch 384/768 PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.
Screenshots of the PatchControl 384 software showing Kir2.1 current traces in response to a voltage step protocol. Measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384) using the whole cell patch methodology and multi-hole chips (4 holes per well), the success rate of valuable data for the analysis was  93%. The IC50 value of Barium was determined as 6.38 µM (Literature: 16.2 µM, Schram et al. Cardiovasc Res. 2003).

KV1.3 - Pharmacology with High Success Rate

SyncroPatch 384PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Evotec.
Shown are screenshots of a pharmacology experiment performed with the SyncroPatch 384PE (a predecessor model of SyncroPatch 384). Recordings from 384 KV1.3 expressing CHO cells were performed simultaneously. Original current traces and the peak current over time are displayed. Data are analysed with DataControl384 full analysis tool. With just a few mouse-clicks normalized concentration response curves can be generated. Here, normalized response and the IC50 of Quinidine is shown. Darkening shades of blue indicate increasing compound concentration.

KV1.5 - Dose response curve of 4-AP

SyncroPatch 384PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.
Screenshots of the PatchControl 384 software showing a dose-response curve of 4-AP on KV1.5 stably transfected cells. Measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384) using multi-hole chips (4 holes per well), the success rate of completed experiments was 100%. The IC50 value of 160 µM corresponds well to literature (IC50 4-AP: 270 µM; Gutman et al., Pharmacological Reviews 57: 473-508, 2005). 

KV4.3 - Pharmacology of Metropolol Tartrate, using the CiPA Protocol

SyncroPatch 384/768 PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.
Screenshots of the PatchControl 384 software showing KV4.3 current traces in response to the CiPA voltage step protocol, measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384) using the whole cell patch methodology and single-hole chips. The IC50 value of Metropolol Tartrate was determined as 128 µM.

KV4.3 - Pharmacology of Quinidine

SyncroPatch 384/768 PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.
Screenshots of the PatchControl 384 software showing KV4.3 current traces in response to the CiPA voltage step protocol. Measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384) using the whole cell patch methodology and multi-hole chips (4 holes per well), the success rate of completed experiments was  95.3%. The IC50 value of Quinidine was determined as 21.2 µM (Literature: 79.3 µM, Crumb et al., J Pharmacol Toxicol Methods. 2016).

KV4.3/KChIP2 - Dose-response curve of Flecanaide

SyncroPatch 384PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.
Screenshots of the PatchControl 384 software showing KV4.3/KChIP2 current traces in response to a voltage step protocol and the corresponding current-voltage relationship plot. Using whole cell mode in combination with multi-hole chips (4 holes per well), stably transfected cells were measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384). The IC50 value of flecainide was determined as 28.3 µM which is in accordance to literature. The success rate of completed experiments was 100%.

KV7.1 (KVLQT) - Dose-response curve

SyncroPatch 384PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.
Screenshots of the PatchControl 384 software showing KV7.1/KCNE (KVLQT/minK) current traces in response to a voltage step protocol and the corresponing current-voltage relationship plot. Using the perforated patch methodology (Escin) in combination with multi-hole chips (4 holes per well), stably transfected cells were measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384). The IC50 value of Chromanol 293B was determined as 3.82 µM. The success rate of completed experiments was 100%. 

NaV1.5 - Current Voltage Relationship

SyncroPatch 384PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by EMD Millipore.
Borosilicate glass chips are used as the patch clamp substrate, ensuring excellent voltage clamp of the cell membrane and high quality seals. Voltage gated channels such as hNaV1.5 (HEK293) have been used to validate the system. This data example shows the current-voltage characteristics and the corresponding raw current traces of a single cell from a recording on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384). The current-voltage plot was fit with a Boltzmann equation revealing a Vhalf of activation of -51 mV for an average of 337 cells.

NaV1.5 - Inactivation Protocol

SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) data and applications:
Cells were kindly provided by EMD Millipore.
Shown are raw current responses of HEK293 cells expressing hNaV1.5 to a double (inactivation) pulse protocol and the corresponding current-voltage plot. The data was fitted with a Boltzmann equation and the Vhalf of inactivation was -84 mV (n = 217).

NaV1.5 - Late Current Analysis using the CiPA Protocol

SyncroPatch 384/768 PE (a predecessor model of the SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.
Screenshots of the PatchControl 384 software showing NaV1.5 current traces in response to the CiPA voltage step protocol, measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384) using whole cell patch clamp methodology and single-hole chips. The NaV1.5 late current was activated by the application of 60 nM ATX-II. The IC50 value of Ranolazine of the late sodium current current was determined as 40.4 µM.

NaV1.5 - Lidocaine Dose Response

SyncroPatch384PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by EMD Millipore
NaV1.5 expressed in HEK293 cells recorded on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384). The concentration response curves for lidocaine block of NaV1.5 were constructed at different holding potential (as indicated) either using a single concentration of compound pera per cell (solid lines) or cumulative concentration response curves (dashed line). The IC50 for lidocaine was shifted by a factor of 35 when holding potential was changed from -120 mV to -80 mV.

NaV1.7 - Accurate Voltage Clamp

SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) data and applications:
Cells were kindly provided by Anaxon AG.
CHO cells expressing NaV1.7 were used on the SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) with a success rate of > 90% for cells which have a seal resistance > 500 MΩ (see inset). A screenshot of the PatchControl 384 software showing current traces in response to a voltage step protocol and the corresponding current-voltage plot.

NaV1.7 - Frequency Dependent Inhibition

SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) data and applications:
Cells were kindly provided by Anaxon.
The frequency dependent inhibition of Tetracaine (5 µM, 15 µM and 50 µM) on NaV1.7 currents were investigated. Measured on the SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) the whole cell patch methodology and single-hole chips were used. The IC50 value determined from the first pulse of the pulse train was calculated as 41.8 µM, from the second pulse as 9.9 µM and from the 10th pulse as 3.0 µM. 

NaV1.7 - Pharmacology of Lidocaine

SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) data and applications:
Cells were kindly provided by Anaxon.
The effect of different concentrations of Lidocaine on NaV1.7 currents were investigated. Measured on the SyncroPatch 384PE the whole cell patch methodology and single-hole chips were used. The success rate of > 90% for cells which had a seal resistance > 500 MΩ was determined. A value of 0.79 was calculated for the z-factor (characterization of HTS screening assay quality).

NaV1.8 - I/V Characteristics

SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) data and applications:
The CHO cells were kindly provided by Charles River.
hNaV1.8 recorded on the SyncroPatch 384PE (a predecessor model of the SyncroPatch 384). Shown are current responses to increasing voltage steps from -60 to +60 mV (left). Current-voltage plot for an average of 380 cells is shown on the right. Shown are mean of peak amplitudes normalized to the maximum of each cell ± S.E.M. The data was fitted using a Boltzmann equation revealing a Vhalf of activation of -2.7 mV (n = 380), in good agreement with the range found in the literature. 

NaV1.8 - State Dependent Inhibition

SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.
State dependent inhibition of tetracaine on NaV1.8 currents was investigated. Measured on the SyncroPatch 384PE (a predecessor model of the SyncroPatch 384), the perforated cell patch methodology (Escin) and multi-hole chips were used and compared to single-whole chips. Using a state dependant pulse protocol, the IC50 value determined from the first pulse (C1) was calculated as 54.3 µM (Hill coefficient = 1.50), and from the second pulse (C2) as 1.27 µM (Hill coefficient = 0.62). 

NaV1.9 - Pharmacology

SyncroPatch 384/768 PE (a predecessor model of the SyncroPatch 384) data and applications:
Cells were kindly provided by Icagen.
Screenshots of the PatchControl 384 software showing hNaV1.9 current traces in response to a voltage step protocol. Measured on the SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) using perforated patch methodology (Escin) and multi-hole chips (4 holes per well), the success rate of valuable data for the analysis was  86%. NaV1.9 is TTX insensitive. The IC50 value of lidocaine of the peak current was determined as 350 µM (Literature: 356 µM), the IC50 value of tetracaine of the peak current was determined as 12.5 µM (Literature: 32 µM).

NMDA NR1/NR2A - Activation and Modulation

SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) data and applications:
Cells were kindly provided by B'Sys.
Here, activation and modulation of the NR1/NR2A subunit containing NMDA receptors are shown. Currents were evoked by application of 10 µM Glutamate in the presence of 10 µM Glycine.
Application of the neurosteroid Pregnenolone sulfate (PS) potentiates the current response induced by Glutamate. The potency of PS was analysed in a single-point screen. The EC50 of PS (39.6 µM), comprising 74% of the cells is nicely corresponding to litereature values (Irwin, et al. Neurosci. Lett. 1992)

P2X2/ P2X3 - Activation by ATP on the SyncroPatch 384PE

SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) data and applications:
Cells were engineered and kindly supplied by Axxam
P2X2/3 recorded from CHO cells on the SyncroPatch 384PE (a predecessor model of the SyncroPatch 384). P2X2/3 was activated by increasing concentrations of ATP with an EC50 = 1.1 µM (n = 191). ATP was applied for approximately 1.5 s using the ‘Ligand Puff’ function of the SyncroPatch 384PE. Average traces and concentration response curve shown for 191 cells. Success rate was 99% for completed experiment (191 wells from a possible 192). Multi-hole (4 holes per well) chips were used.

P2X2/ P2X3 - Block by suramin on the SyncroPatch 384PE

SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) data and applications: 
Cells were engineered and kindly supplied by Axxam
P2X2/3 recorded from CHO cells on the SyncroPatch 384PE (a predecessor model of the SyncroPatch 384). P2X2/3 was activated by 10 µM ATP and blocked by increasing concentrations of suramin with an IC50 = 17.7 ± 0.9 µM (n = 372). Average traces and concentration response curve shown for 372 cells. Success rate was 97% for completed experiment (372 wells from a possible 384). Multi-hole (4 holes per well) chips were used.

P2X2/ P2X3 - High throughput screening on the SyncroPatch 384PE

SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) data and applications: 
Cells were engineered and kindly supplied by Axxam
Inhibition of P2X2/3 by suramin. Suramin was pre-incubated at each concentration for 3 - 5 mins followed by co-application with 10 µM ATP. All 3 concentrations of suramin were applied to each well and a cumulative concentration response curve was constructed for n = 372 wells out of a possible 384. Success rate was 97% for completed experiments.

Piezo1 in Neuro2A cells - activation by Yoda1

SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) data and applications:

Piezo1 channels endogenously expressed in Neuro2A cells were investigated on the SyncroPatch 384PE (a predecessor model of the SyncroPatch 384). Screenshot of the PatchControl 384 software during an experiment. B Statistical analysis of the currents at -100mV (left) and at 80 mV (right). 140 out of 384 Neuro2A cells (37%) passed the quality criteria and 85 cells (60% of the valid cells) were considered as Yoda1 responders.

Data from Rotordam et al, 2019.


Piezo1 in red blood cells - activation by Yoda1

SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) data and applications:

Current response of Piezo1 activated by Yoda1 in patient cells with the novel PIEZO1 mutation (R2110W) compared to healthy red blood cells (RBCs). Shown are raw data traces (top) and statistical analysis of all measured cells, independent of their response to Yoda1 (bottom).

Data from Rotordam et al, 2019.


Piezo1 in red blood cells - Hereditary Xerocytosis

SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) data and applications:

Whole-cell recordings of ion currents from RBCs of healthy donors and Hereditary Xerocytosis patients. Different mutations in the PIEZO1 gene were compared with controls. Aa The P50.2 mutation resulted in current conductance that was unchanged compared with transport controls, but showed increased conductance compared with general controls (Ab). The mutation P52.1 showed decreased conductance compared with transportation controls (Ba) and general controls (Bb).

Data from Petkova-Kirova et al, 2019.


TRPA1 - Activation by carvacrol

SyncroPatch 384i (a predecessor model of the SyncroPatch 384) data and applications: 
Cells were kindly provided by AcCELLerate.
Activation of TRPA1 expressed in CHO cell on the SyncroPatch 384i (a predecessor model of the SyncroPatch 384). A Screenshot of the PatchControl 384 software depicts raw data traces of TRPA1- expressing CHO cells as recorded on one NPC-384 patch clamp chip (4 holes). A single concentration of carvacrol was added to each well and the concentration response curve calculated across the plate. B Average CRC (left) for carvacrol for n = 355 wells and average traces (right). TRPA1 was robustly activated by carvacrol. At higher concentrations (above 400 µM) desensitization of the channel was observed (tachyphylaxis) which resulted in smaller amplitudes upon repeated application.

TRPA1 - Inhibition by A967079 and AMG0902

SyncroPatch 384i (a predecessor model of the SyncroPatch 384) data and applications: 
Cells were kindly provided by AcCELLerate.
Inhibition of TRPA1 by A967079 and AMG0902. A TRPA1-mediated responses were blocked by A967079 in a concentration-dependent manner with an IC50 value (12.4 nM) in good agreement with the literature value of 50 nM. B TRPA1-mediated responses were blocked by AMG0902 in a concentration-dependent manner with an IC50 value of 48 nM (n = 349) in good agreement with the literature value of 68 nM.

TRPM8 and Temperature Control

SyncroPatch 384 data and applications:
Cells were kindly provided by Charles River Chantest
At RT, the TRPM8 current was activated using increasing concentrations of Menthol (left). Measurement site, cells and solutions can be accurately temperature controlled – in the presence of temperatures 38°C, 25°C, 18°C and 12 °C the current size of temperature regulated TRPM8 changes accordingly (right). TRPM8 is activated at temperatures < 25°C.



インタビュー&ケーススタディー

Dr. David Dalrymple - Statement about the SyncroPatch 384PE

“As a leading ion channel contract research organization running one of the most comprehensive ranges of ion channel assays, SB Drug Discovery has been impressed with the flexibility and reliability of the SyncroPatch 384PE (a predecessor model of SyncroPatch 384i), enabling development of a range of varied and complex ion channel assay for both high throughput screening and hit-to-lead profiling purposes. The SyncroPatch has proven to be a crucial addition to SB’s ion channel capabilities and in partnership with expert advice from Nanion’s support team has enabled SB to advance its ion channel capabilities to the forefront of ion channel drug discovery research..“
 
Dr. David Dalrymple
Business Development Director at SB Drug Discovery

Prof. Al George - A New Era is Emerging for Ion Channel and Channelopathy Research

“... the SyncroPatch has revolutionized our ability to determine the functional consequences of hundreds of human ion channel variants, which could be considered one of the most significant recent advances in channelopathy research.”


Prof. Dr. Al George
Professor and Chair of Pharmacology at Northwestern University Feinberg School of Medicine, Chicago, IL, USA

Download the full customer case study here:

 

SyncroPatch 384i (a successor of SyncroPatch 384PE) Customer Case Study: (3.1 MB)

Prof. Dr. Al George, Professor and Chair of Pharmacology at Northwestern University Feinberg School of Medicine, Chicago, IL, USA


Prof. Al George - Statement about the SyncroPatch 384PE

“We are extraordinarily excited about installing the first SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) in an academic setting in North America. The enormous throughput, intuitive software and robust liquid handling capabilities along with superior seal quality, stability and high success rates convinced us to purchase the instrument. The SyncroPatch 384PE will enable us to perform detailed high throughput analysis of genetic variants in human ion channels at a previously unobtainable scale, and will form the cornerstone of a new HTS facility we are building. We also look forward to upgrading to 768 wells in the near future.“

Dr. Al George
Professor and Chair of Pharmacology at Northwestern University Feinberg School of Medicine, Chicago, IL, USA


Prof. Jamie Vandenberg - Statement about SyncroPatch Technology

“In my lab we use the SyncroPatch 384PE, as it allows us to reliably screen KCNH2 mutations, the gene encoding hERG K+ channels, in high-throughput. This is a very fast process and it enables us to screen large mutation libraries. Recently, we have validated a high-throughput functional phenotyping assay capable of distinguishing benign KCNH2 variants from those that have a dominant negative effect, using the SyncroPatch 384PE. We are very happy with this result and plan to expand assay development to other ion channel genes.”

Prof. Dr. Jamie Vandenberg, co-deputy director and head of cardiac electrophysiology 
The Victor Chang Cardiac Research Institute, Australia

Nina Braun - Statement about the SyncroPatch 384i

“I measured CRCs for activation and steady-state desensitization, as well as peptide modulation of the channel, and got fantastic support from Søren Friis both with technical questions and assay design .The SyncroPatch and its software are easy to use and allow for versatile assay design. The team at Nanion goes out of their way to help with all questions and requests that come up, and they host fantastic user meetings for idea exchange. After four years using the SyncroPatch, I can highly recommend it.”“

Nina Braun 
University of Copenhagen




チュートリアル

Basic principles of external solution exchange and compound addition

Also applies to the SyncroPatch 384




ウェビナー&動画


14.10.2020 | Webinar: Development and validation of ASIC1a ligand-gated ion channel drug discovery assays on automated patch clamp platforms

SyncroPatch 384i and Patchliner Webinar
Date: October 14. 2020
Speakers: Dr. Marc Rogers (Metrion Biosciences; U.K.)
This is an on-demand webinar from Nan]i[on and Friends 2020.

15.10.2020 | Webinar: Turning Cells into Reagents

SyncroPatch 384i Webinar
Date: October 15. 2020
Speakers:Dr. Oliver Wehmeier (acCELLerate GmbH) and Dr. Tim Strassmaier (Nanion Technologies)
This is an on-demand webinar from Nan]i[on and Friends 2020.

15.10.2020 | Webinar: ICH S7B best practices considerations - New Q&As and Benchmarking best practices

Patchliner and SyncroPatch 384i Webinar
Date: October 15. 2020
Speakers: Dr. Sonja Stoelzle-Feix (Nanion Technologies)
This is an on-demand webinar from Nan]i[on and Friends 2020.

15.10.2020 | Webinar: Benchmarking best practices and calibration standards for HTS hERG recordings for improved proarrhythmic assessment

Patchliner and SyncroPatch 384i Webinar
Date: October 15. 2020

Speakers:Dr. Alison Obergrussberger (Nanion Technologies)

This is an on-demand webinar from Nan]i[on and Friends 2020.


16.10.2020 | Webinar: Emerging Role of LRRC8 Volume-Regulated Anion Channels in the Skin

SyncroPatch 384i Webinar
Date: October 16. 2020
Speakers:
Dr. Torsten Fauth (BRAIN AG)
Dr. Oliver Rauh (Technical University of Darmstadt)
Giustina Rotordam (Nanion Technologies)
 
This is an on-demand webinar from Nan]i[on and Friends 2020.

2020 - What is the Syncropatch 384PE and what are the benefits of the Syncropatch 384PE?

SyncroPatch 384PE product video
The Victor Chang Cardiac Research Institute's Innovation Centre is one of the only two centres in Australia that houses the Syncropatch 384PE. The Syncropatch 384PE enables high throughput quantification of electrical signals in cells. In this video, Dr Jeffrey McArthur explains how it works, the advantages of using the Syncropatch384PE to fast track your experiments and how it can benefit your research.

2019 - SyncroPatch 384i Product Video

SyncroPatch 384i product video
 

The SyncroPatch 384i – is a giga-ohm seal HTS automated patch clamp platform based on the newly introduced and state-of-the art Biomek i5 liquid handler.

It provides effortless ion channel screening coupled with unmatched flexibility, ease-of-use and reliability. The SyncroPatch 384i builds on the success of the SyncroPatch 384PE, which has been globally established as the preferred automated patch clamp workhorse in Pharma, Biotech, CRO and academia.


2018 - HTS Phase I study: an update on progress of the CiPA Ion Channel Work Stream using the SyncroPatch 384PE and Patchliner

SyncroPatch 384PE (a predecessor model of the SyncroPatch 384i), Patchliner and CardioExcyte 96 Oral Presentation
Presenter: 
Tim Strassmaier, Nanion Technologies Inc. USA
Source:
Webinar: "CiPA study: Bridging ion channel and myocyte data", September 12, 2018

2018 - Biophysical and Pharmacological Characterization of Voltage-Gated Sodium Channels Involved in Pain Pathways

SyncroPatch 384PE (a predecessor model of the SyncroPatch 384i) Oral Presentation Video
Presenter: 
Dr. Markus Rapedius, Senior Scientist, Nanion Technologies

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