• Patchliner

    兼顾科研与工业的全自动膜片钳系统
  • Patchliner

    自主生产与质量控制的耗材
  • Patchliner

    拥有超过10年实验设计与支持的经验
  • Patchliner

    拥有所有手动膜片钳的特征与优点
  • Patchliner

    全球已经安装超过100台Patchliner系统

Patchliner - 兼顾科研与药物研发

Patchliner全自动膜片钳最多可同时记录8个细胞,具有极大的实验自由度和极高的数据质量。Patchliner是市场上最通用的膜片钳系统之一。Patchliner具有以下特点:

硬件:

  • 可对细胞外液与内液进行灌流
  • 可无限次数给药(由于持续对废液移除)
  • 温度控制 (见Applications)
  • 可制冷的细胞池
  • 支持电压钳 & 电流钳记录 + 动态钳制技术
  • 有4通道与8通道两个配置可供选择
  • 有自产的单孔 & 多孔芯片
  • 串联电阻Rseries补偿

应用:

  • 电压与配体门控离子通道
  • 温度激活通道
  • 在生理温度下记录
  • 全细胞l & 穿孔膜片钳
  • 细胞系、原代细胞与干细胞
  • 为CiPA而验证的系统
  • 极少的细胞消耗
  • 易用 & 定制的分析工具

Patchliner是一款多功能且强大的膜片钳系统,是基础研究与离子通道生物物理学与通道活动机制研究的理想工具。 可进行许多复杂的实验,如热激活TRP 通道, 通过 内液交换 激活 Ca2+-激活通道与通过快速外液灌流实现对诸如 nAChα7配体门控离子通道进行短时间的药物暴露操作。 such as Patchliner也是进行常规试验的卓越的工具,例如CiPA规定的对ERG和其他心脏离子通道的安全筛选等。

自2006年发布以来, 在全球已经安装了超过100台设备,包括学术机构 (46%), 制药企业(34%)与CRO公司 (20%)。Patchliner由于其稳定的高封接成功率 (>80% GΩ级封接), 极少的细胞消耗和被证明能在原代细胞与干细胞上使用等优势点而被广泛赞誉。

我们专业的电生理团队与工程师们持续进行内部的试验研发, 软件与硬件的升级、提供快速与定制的溶液以满足实验需求。

更多信息:

Patchliner 软件

PatchControl HT

screenshot pl sw

PatchControl HT is the software for the Patchliner. The vast experimental freedom of the Patchliner platform is due to the flexibility of the software.

PatchControl HT is a graphical user interface allowing straightforward and easy programming of protocols either by using pre-programmed modules, or by setting up customized protocols based on the specific user requirements.

Because of its inherent logical programming capabilities, experimental redundancy is reduced to a minimum, thus maximizing the data throughput.

PatchControl HT not only allows the running of experiments from start to finish, it also supports changes on-the-fly. This latter feature speeds up the assay development process immensely. It also makes the Patchliner a valuable tool for research.

In addition, PatchControl HT is compatible with various document and database formats which makes compound loading and data analysis faster and easier when screening a large number of compounds.


分析软件

hERG Screenshot Data Analysis Tool 1 700

Data can easily be analyzed using Nanion's Data Analysis Package, a very efficient and convenient data analysis tool. With a few mouse clicks raw data is loaded, displayed, analyzed and pooled, reducing the effort to a minimum.

Display of raw current traces at this point allows judgement of the quality of the data recorded from each cell. Individual cells can be easily excluded from the data set and averages recalculated.

Current Voltage relationship plots as well as IC50 values are automatically calculated and displayed.

耗材

NPC-16芯片

application16 chip 4

The NPC-16 chip is a proprietary innovative product by Nanion Technologies, developed for the Patchliner. It is produced and quality-assured in-house at Nanion headquarters and shipped from Munich to our international customers. Different types of NPC-16 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 cartridge forming the microfludic channels through which the solutions are delivered. The design of the chip allows as well an internal solution exchange during an experiment.


Features

Each NPC-16 chip contains 16 recording chambers.
Depending on the Patchliner version (Quattro, Octo) 4 or 8 of these sites are used at a time. Three chips can be loaded onto the Patchliner. 48 recordings can be performed without user intervention.
Additionally, the number of exchanges of either the internal or the external solution is unlimited.


Available chip types
  • "NPC-16, 1x, medium resistance": One hole well, 1.8 - 3 MOhm (Order # 071102)
  • "NPC-16, 4x, medium resistance": Four hole wells, 1.8 - 3 MOhm (Order # 071401)
  • "NPC-16, 1x, low resistance": One hole wells, 1.5 - 2 MOhm (Order # 071103)
  • "NPC-16, 1x, high resistance": One hole wells, 3 - 5 MOhm (Order # 071101)
  • "NPC-16, 1x, ultra-high resistance": One hole wells, 5 -8 MOhm (Order # 071104)

Other sizes are available upon request!

溶液

Buffers and Solutions for the Patchliner

The buffers and solutions for the Patchliner are produced by an external partner, quality-assured in-house at Nanion headquarters and shipped from Munich to our international customers.


Available buffers and solutions
  • "Reagent Kit Patchliner (1 month) standard": 20x 50 mL External Solution; 5x 50 mL Internal Solution (Cs), 5x 50 mL Internal Solution (K) (Order # 081007)
  • "Reagent Kit Patchliner (1 month) to be specified by the customer": (Order # 081017)
  • "Reagent Kit Patchliner (3 months) standard": 60x 50 mL External Solution; 15x 50 mL Internal Solution (Cs), 15x 50 mL Internal Solution (K) (Order # 081008)
  • "Reagent Kit Patchliner (3 months) to be specified by the customer": (Order # 081018)

数据与应用

Acetylcholine Receptor Alpha 7 - Enhancement by NS1738

nACha7R PL NS1738 DRCicon pl   Patchliner data and applications:
Cells were kindly supplied by Galantos Pharma GmbH.

Concentration response curve for NS1738 co-applied with 300 μM ACh revealed an EC50 = 2.6 ± 1.1 μM (n = 4). This is in excellent agreement with the literature (Timmermann et al, 2007, JPET 323: 294–307).

Acetylcholine Receptor Alpha 7 - Enhancement of cabamoylcholine and epidatidine responses by PNU120596

ACha7 PNU Sheffelicon pl   Patchliner data and applications:
Cells were kindly supplied by Galantos Pharma GmbH.

Representative current records of carbamoylcholine- and epibatidine-induced nAChR activation in the absence and presence of PNU-120596. Horizontal bars indicate exposure time (233 ms) of compound(s) to the cell. Data from Scheffel et al, 2018.

Acetylcholine Receptor Alpha 7 - Enhancement of nicotine and acetylcholine responses by PNU120596

nACha7R PL PNU120596 scheffelicon pl   Patchliner data and applications:
Cells were kindly supplied by Galantos Pharma GmbH.

Representative current records of hα7-nAChR responses induced by nicotine and acetylcholine (ACh) (50, 100, 500 μM) in the absence (top) and presence of PNU-120596 (5, 10, 50 μM) (bottom). Horizontal bars indicate exposure time (233 ms) of compound(s). Data from Scheffel et al, 2018.

AMPA Receptor (GluA2) - Inhibition by CNQX

icon pl  GluA2_CNQX_SB_PLPatchliner data and applications:
Cells were kindly provided by SB Drug Discovery.

The AMPA receptor (GluA2) was blocked by CNQX on the Patchliner. CNQX was pre-incubated and then co-applied with glutamate. CNQX blocked the GluA2-mediated response in a concentration dependent manner and the potency was dependent on glutamate concentration (left). Exemplar GluA2-mediated responses are shown on the right activated by 100 µM glutamate and inhibited by increasing concentrations of CNQX.

hERG - Pharmacology of Cisapride, using the CiPA protocol

icon pl   CiPA PL hERG CisapridePatchliner data and applications:
Cells were kindly provided by Charles River.

The effect of cisapride on hERG currents was investigated, using the CiPA voltage step protocol. Measured on the Patchliner the perforated patch methodology (Escin) and multi-hole chips (4 holes per well) were used. The IC50 value of Cisapride was determined as 112 nM.

Cardiac Ion Channels - Pharmacology of Vandetanib

CiPA PE CE Pharmacology VandetanibIcon CE    CardioExcyte 96 and   icon pl   Patchliner 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 Vandetanib on hERG, NaV1.5, CaV1.2 and KV4.3. The compound induced arrhythmia when iPSC-CM were exposed to a minimum concentration of 1 µM. Arrhytmic events were both detected in field potential recordings as well as in the impedance based contractility.

GABAA Receptor - Currents in iCell Neurons

GABA Bicuculline neurons

icon pl   Patchliner data and applications:
Cells were kindly provided by Cellular Dynamics.

Activation of GABAA receptor currents by 30 μM GABA and partial block of the current response by 1 μM bicuculline. Bicuculline was pre-applied for at least 30 s before co-application with GABA (30 μM). Approximately 50% of the current was blocked by 1 μM bicuculline.

Neurons (iPSC-derived) - Voltage Gated Potassium Currents

KvIV Neurons Figure3 icon pl   Patchliner data and applications:
Cells were kindly provided by Cellular Dynamics.

A Voltage-gated K+ current recorded in iCell® neurons. Current responses to a voltage step protocol. An outward K+ current can be seen in this cell.
B Corresponding IV plot from an average of 7 cells. 

Neurons (iPSC-derived) - TXX Block of Sodium Currents

TTX Neurons Figure2icon pl   Patchliner data and applications:
Cells were kindly provided by Cellular Dynamics.

A Block of Na+ current by increasing concentrations of TTX. Current was blocked by low nM concentrations of TTX indicating a TTX sensitive Na+ channel type expressed in this cell.
B Concentration response curve for TTX inhibition, IC50 = 4.9 nM (n =1).

Neurons (iPSC-derived) - Voltage-Gated Sodium Currents

NaIV Neurons Figure1icon pl   Patchliner data and applications:
Cells were kindly provided by Cellular Dynamics.

Voltage-gated Na+ current recorded in iCell® Neurons. Current responses to a voltage step protocol.
A large inward Na+ current can be seen in this cell with a K+ outward current present at positive voltages.
B Normalised IV plot from an average of 4 cells.

NaV1.8 - Voltage Dependent Block by Tetracaine

IC50 TetracaineOverlayicon pl   Patchliner data and applications:
Cells were kindly provided by Millipore.

Recordings were made on the Pachliner. The potency of tetracaine was affected by holding potential, becoming less potent with a more negative holding potential. Average concentration response curve for tetracaine, IC50 = 35 ± 8 μM (n = 3) for Vhold - 90 mV and 74 ± 15 μM (n = 4) for Vhold - 120 mV.

 

NaV1.8 - Tetracaine Pharmacology

ND723 Nav18 Tet Figureicon pl   Patchliner data and applications:
Cells were kindly provided by Millipore.

A Raw traces from an exemplar cell recorded on the Patchliner showing inhibition of current by increasing concentrations of tetracaine. Shown are current responses to a single step protocol to 20 mV for 25 ms from a holding potential of -90 mV. Current amplitude was completely recovered upon washout of tetracaine (red trace).
B Timeplot of the experiment.

NaV1.8 - Current-Voltage Relationship

ND723 PL Nav18 IV Figureicon pl   Patchliner data and applications:
Cells were kindly provided by Millipore.

A Raw traces from an exemplar cell recorded on the Patchliner. Shown are current responses to increasing voltage steps from -80 to +60 mV.
B Average current-voltage plot, Vhalf of activation was -9 mV (n = 19).
C Average inactivation plot, Vhalf of inactivation was -24 mV (n = 4). NaV1.8 currents started to activate at about -40 mV, peak response was elicited at around 20 mV.

CaV3.2 - Mibefradil Antagonism

Cav32 Mibefradil

icon pl   Patchliner data and applications:
Cells were kindly provided by Millipore.

Dose dependent block by Mibefradil on current traces from an individual cell expressing CaV3.2. Data was averaged and fitted to the Hill equation

 

CaV3.2 - Inactivation

Cav32 inactivation

icon pl   Patchliner data and applications:
Cells were kindly provided by Millipore.

Current responses of a double pulse protocol with varying test potentials between the pulses (5 s) was used to determine the half inactivating potential. Peak current responses to the second pulse are expressed relative to the response to the first pulse. Both curves in Figure 5 were fitted to the Boltzmann equation and revealed a half-inactivating potential of -65 mV and a half-activating potential of -33 mV.

CaV3.2 - Current-to-Voltage Relationship

IV Cav32

icon pl   Patchliner data and applications:
Cells were kindly provided by Millipore

Representative current responses of an individual cell expressing Ca 2.2 to a standard voltage protocol. The average mean current at -20 mV of all recorded cells was -785 ± 110 pA (n = 12).

CaV2.2 - Cadmium Block

Cd DR

icon pl   Patchliner data and applications:
Cells were kindly provided by Millipore.

The image shows current response of an individual cell in the presence of increasing cadmium concentrations. The IC50 was calculated from the Hill fit to be 3.6 ± 0.4 μM (n = 5).

 

CaV2.2 - Current Voltage Relationship

Cav22 DR

icon pl   Patchliner data and applications:
Cells were kindly provided by Millipore

Representative current responses of an individual cell expressing CaV2.2 to a I/V voltage protocol. The average peak current at 30 mV of all recorded cells was -698 ± 115 pA (n = 6).

 

TRPM3 - Positive modulation

TRPM3 PS C Dataicon pl   Patchliner data and applications: 
Cells were kindly provided by Prof. Thomas Voets, KU Leuven, Belgium.

A Current responses to a voltage ramp protocol from -150 mV to 150 mV over 200 ms in response to 100 μM pregnenolone sulphate (PS) and in combination with increasing concentrations of compound C. B Current responses elicited by PS, enhancement by 30 μM compound C and block of the current by co-application of antagonist (10 μM).

Cardiac Action Potentials - Automated recordings from iCells

0AP Patchliner CDI

icon pl   Patchliner data and applications:
The stem cell-derived cardiomyocytes (iCell) were kindly supplied by Cellular Dynamics.

In this example both Na+ and Ca2+ mediate the action potential. When nifedipine is applied in the current clamp mode, the action potential is shortened significantly due to block of the calcium channels.

 

Acetylcholine Receptor Alpha 7 - Stable nicotine responses

icon pl   Patchliner data and applications: nAChRa7 repetitive PL
Cells were kindly supplied by Galantos Pharma GmbH.

Stable whole-cell current amplitudes were obtained by repeated 100 mM nicotine stimulation of HEK293 cells expressing human nACha7 receptors. The stacked application protocol was used. 

Acetylcholine Receptor Alpha 7 - Activation and Dose Response Curve

PL Nica7 DRicon pl   Patchliner data and applications:
Cells were kindly supplied by Galantos Pharma GmbH.

Complete nicotine dose response curves were obtained by applying increasing concentrations of nicotine to a HEK293 cell expressing human nicotinic α7 acetyl choline receptors. The stacked application protocol was used.

TRPV3 - Temperature Activation

icon pl   Patchliner data and applications:TRPV3 Patchliner
Cells were kindly supplied by Millipore.

Recordings from a HEK cell expressing TRPV3 were made with the Patchliner showing activation by heat. External solution was heated inside the Patchliner pipette to the temperature shown and applied to the cells. TRPV3 was activated at temperatures ≥ 38°C.

 

hERG - Application of "Sticky Compounds"

application16 herg 2

icon pl   Patchliner data and applications:
hERG expressing HEK293 cells were kindly provided by Cytomyx/Millipore.

Even sticky compounds pose no problem for the Patchliner. IC50 measurements of well known sticky substances were determined on the Patchliner: Terfenadine IC50 = 11.0 ± 3 nM, Flunarizine IC50 163.7 ± 19 nM and Cisapride IC50 8.9 ± 3 nM.

Gramicidin - Bilayer Recordings

p37 2 BilayRec

icon pl   Patchliner data and applications:

With suction the GUVs are attracted to the aperture. As soon as one GUV hits the glass substrate, it bursts and forms a bilayer across the aperture. Shown are single channel recordings from gramicidin which was incorprated into the bilayer after its formation. Traces were recorded in 100 mM HCl at −100 mV.

GABAA receptors - Investigation of Modulators

p37 1 modulatorsicon pl   Patchliner data and applications:
Cells were kindly provided by AstraZeneca.

The top images show dose dependent block of GABAA currents by bicuculline. The IC50 was determined as 1.2 ± 0.2 μM (n=11). The lower graph shows the positive modulation of glycine activation of hGlyRα1. Here, six co-applications of 20 μM glycine and increasing concentrations of a positive modulator are shown.

Glycine Receptor (GlyRa1) - Accurate Pharmacology

p36 4 glycineicon pl   Patchliner data and applications: 
Cells were kindly provided by AstraZeneca.

Two different application protocols were used to study the effect of exposure time on glycine receptor pharmacology for cells expressing hGlyRα1. As shown from the raw data traces and corresponding Hill plots, the highest concentration, 3 mM glycine, did not elicit the maximum peak response during long exposures (22 s), in contrast to stacked applications (1 s).

GABAA Receptor - Stacked Application Technology

p36 3 timedIntervicon pl   Patchliner data and applications:

Ligand gated ion channels often display receptor desensitization. A method was developed to minimize ligand exposure times and intervals between ligand exposures. The pipette first aspirates buffer, then compound. When expelling this stack, the cell is first exposed to ligand and then buffer. Exposure times as low as 400 ms are possible with this method. A GABA dose response curve, aquired in this manner, is shown on the left.

TRPV1 - Transient Heat Activation

p36 2 TRPV1icon pl   Patchliner data and applications:

The current responses of a CHO cell expressing TRPV1 (ramp -100 mV to +100 mV) at increasing temperatures is shown. The ET50 value was determined as 64°C. Challenge of the same cell with capsaicin (1 μM) and temperature (70°C) allows comparison of the responses. IC50s for ruthenium red block of capsaicin- and heat-responses were determined as 1.6 ± 0.2 μM (n = 3) and 7.4 ± 1.3 μM (n = 3), respectively.

KV1.3 - Reproducible Compound Application

icon pl   Patchliner data and applications:p36 1 reproducRec

Application of 5 μM quinidine leads to about 50 % block of the KV1.3 currents (blue). After washout, the current is fully recovered (grey). The lower graph shows corresponding Imax (+40 mV) in the absence and presence of 5 μM quinidine, for two different cells with eight consecutive application and washout steps. The recording lasted over 40 minutes!

KV1.3 - Internal Solution Exchange during Recording

p35 4 ExchInternSolicon pl   Patchliner data and applications:

A unique feature of the Patchliner is its ability to exchange the internal solution during the recording. The figure shows recordings of KV1.3 from two Jurkat cells (simultaneoulsy recorded) in the presence of control internal solution, after the exchange of the internal solution with a Cs+ solution, and subsequent washout (left to right).

Astrocytes - Internal Perfusion

p35 3 ratAstrocytes 2icon pl   Patchliner data and applications:
Data courtesy of C. Peers, University of Leeds, Leeds, UK.

Whole cell currents from rat cortical astrocytes (primary culture) were evoked by 500 ms long depolarizing voltage steps (-100 mV to +40 mV). Currents were blocked by administration of internal Cs+, and recovered when switching back to Cs+-free internal solution. Averaged data are presented as mean ± S.E.M. (n=35). For more information, see Nature 254, 4 (2), 2009.

NaV1.5 - Stable Access Resistance

p35 2 VoltContricon pl   Patchliner data and applications: 
Cells were kindly provided by Millipore.

The I/V-characteristics of NaV1.5 currents (HEK293) are shown together with the repeated dose dependent block by TTX (lower panel). Five concentrations of TTX (0.3, 1, 3, 10, 30 μM) were applied, followed by washout with antagonist-free buffer and re-application of the same TTX concentrations. 

Cardiac Ion Channels - Pharmacology of Sodium Channels

p35 1 CorAticon pl   Patchliner data and applications:
Cells (Cor.AT) were kindly provided by Axiogenesis.

The pharmacology of dibucaine was investigated by the application of 0.3, 1, 3, 10 μM in the presence of 10 μM nifedipine (L-type Ca2+-current blocker). Two control additions of nifedipine (10 μM) were made before the addition of increasing concentrations of dibucaine. The IC50 value was determined as 355 ± 40 nM (n=3), which is in accordance with the literature.

Cardiac Ion Channels - Recordings from SC-Derived Cardiomyocytes

p34 4 actPot

icon pl   Patchliner data and applications:
Cells were kindly provided by Ncardia.

The left picture shows a typical action potential from Cor.At® cardiomyocytes. Whole cell currents recorded in the voltage clamp mode reveal cardiomyocyte-typical ion channels (right). The traces represent mERG-, L-type Ca2+- (blue, block by 50 μM nifedipine), Na+- and K+-currents (from top left to bottom right).

 

hERG - Efficient Screening

p34 2 hERG II Sum

icon pl   Patchliner data and applications:
Cells were kindly provided by Cytomyx/Millipore.

The effects of six different blockers (terfenadine, cisapride, E4031, astemizole, propafenone, quinidine) on hERG currents (HEK293 cells) were investigated. Expected IC50 values for the different compounds were obtained. In two days, 119 full dose response curves were collected by a single person. Data was analyzed using Nanion’s Data Analysis Package, a very efficient and convenient data analysis tool!

Cardiac Action Potentials - From SC-Derived Cardiomyocytes

AN Patchliner CorAtCardiomyocytes 1

 icon pl   Patchliner data and applications:
Cells were kindly provided by Ncardia.

Action potentials recorded from stem-cell derived cardiomyocyetes (Cor.At® cardiomyocytes). Action potentials are triggered by small current pulses. Effects of quinidine and lidocaine on the action potentials are shown.

  

hERG - Simple Data Analysis

hERG Screenshot Data Analysis Tool 1 700

icon pl   Patchliner data and applications:

With our analysis tools, especially programmed routines in Igor make dose response curves, raw data and current time courses easily accessible. Also, creating average dose response curves over multiple experiments - even conducted on different days - remains easy.

hERG - Block at Physiological Temperature

AppNote hERG TEmp

icon pl   Patchliner data and applications:
Cells were kindly provided by Cytomyx/Millipore, UK.

The effects of erythromycin on hERG currents were tested at different temperatures. Erythromycin has been shown to block hERG channels at physiological temperature with an IC50 of approx. 40 µM. However, at RT erythromycin is much less potent. For more details on these experiments please refer to the Application Note

 

AMPA Receptor (GluA2) - Fast Activation

GluR2 CRC current time course

icon pl   Patchliner data and applications:
Cells were kindly provided by University of Sussex.

Shown is concentration dependent activation of GluA2 receptors (known as AMPA receptors) by 10 μM, 30 μM, 100 μM, 300 µM and 1 mM Na-Glutamate from a GluA2 expressing HEK293 cell. The rising phase is enlarged in the right graph.

Acetylcholine Receptor Alpha 3 Beta 4 - Concentration Response Curve to Nicotine

nAChRa3b4_nic_DRC_PL

icon pl  Patchliner data and applications:

Shown are the a raw current responses of a HEK293 cell expressing AChR (α3β4) to increasing concentrations of nicotine. Solutions were  stacked (layered) in the pipette to achieve brief exposure times.

 

Erythrocytes - Whole Cell Recordings

PatchlinerRBC

icon pl   Patchliner data and applications:
Cells were kindly donated by Dr. Andrea Brüggemann.

Whole cell current recordings from erythrocytes recorded on an eight-channel Patchliner.

 

 

 

 

Erythrocytes - Single Channel Recordings

application16 erythro 1

icon pl   Patchliner data and applications:

The membrane of erythrocytes contains different ion channels like Ca2+-activated K+ channels, or the volume-sensitive Na+/K+ pump. Studies also revealed the participation of a Ca2+-permeable non-selective cation channel in the regulation of erythrocyte 'apoptosis'. Shown are single channel fluctuations as recorded from an erythrocyte in the cell attached configuration on the Patchliner.

CFTR - Regulation

CFTR

icon pl   Patchliner data and applications:

The Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is activated by forskolin. The upper graph shows the timecourse of currents recorded at +95 mV. The bar above the data indicates the time of compound application. Arrows indicate data from which time points were averaged in the lower figure (n = 3).

You can download the full report here.

 

 

Kir channels - Rapid external solution exchange study in RBL cells

icon pl   Patchliner data and applications:application16 kir2 2 small

The basophilic leucaemia cells (RBL) exhibit an inwardly rectifying potassium current, Kir. Changing the external K+ concentration (here between 4.5 mM and 143 mM) leads to a change of the current amplitude of the inward current (holding: -100 mV). This gives us a convenient tool to study the speed of the external solution exchange which was determined as ~50 ms.

hERG - Stable Recordings

application16 herg 1 small

icon pl   Patchliner data and applications:
Cells were kindly provided by Cytomyx/Millipore, UK.

A series of drug concentrations can be applied to each cell. The top figures show the original traces and the corresponding average dose-response curve. Five concentrations of Quinidine (0.1, 0.3, 1, 3 and 10 μM) have been applied.

The lower figure shows the corresponding Imax (-40 mV) including a wash out step and an additional application of the blocker to demonstrate the stability of whole cell recordings.

 

NaV1.5 - Lidocaine Block

application hnav15 3 small

icon pl   Patchliner data and applications:
Cells were kindly provided by Cytomyx/Millipore.

Full dose response curves at different holding potentials were recorded for each cell (hNav1.5 in HEK293). Currents were elicited by a 10 ms voltage step to 0 mV. Plotted are average peak currents as a function of holding potential and lidocaine concentration. 

TRP Channels - Currents in Smooth Muscle Cells

smoothMuscleicon pl   Patchliner data and applications:
Data are taken from Milligan C.J. et al., Nature Protocols, 2009, 4(2), 244-255.

Left: Illustrative time series showing currents at +80 mV and −80 mV in a smooth muscle cell exposed to extracellular Gd3+ (100 μM) and then 2-APB (75 μM). Right: Membrane resistance (Rm), series resistance (RS) and membrane capacitance (Cm) values for successful recordings from smooth muscle cells.

P2X7 - Application of BzATP

P2X7icon pl   Patchliner data and applications:
Data are taken from Milligan C.J. et al., Nature Protocols, 2009, 4(2), 244-255.

BzATP concentration response curve obtained from wild-type P2X7 receptors. Data are fite to the Hill equation with an EC50 of 69.7 ± 7.5 μM. The inset show representative currents evoked by BzATP (3 − 100 μM). Holding potential was −60 mV.

TRPA1 - Current Time Course

TRPA1 Patchlinericon pl   Patchliner data and applications:
Cells were kindly supplied by Millipore.

Shown is the time course of the current measured at +95 mV from an individual cell. The legend indicates the time periods in which the activator AITC was applied. Here we demonstrate the reproducibility of the current responses for TRPA1.

TRPA1 - Current Traces

TRPA1 Screenshoticon pl   Patchliner data and applications: 
Cells were kindly supplied by Millipore.

Shown is a screenshot from a recording on a 4-channel Patchliner from HEK293 cells expressing TRPA1. Currents were activated by ca. 20 s application of 3 μM AITC followed by wash out. On the left raw whole cell currents as responses to 0.2 s voltage ramps (−100 mV to +100 mV) which were applied every 10 s are shown. On the right currents at +95 mV are plotted against time.

TRPC5 - Block by Gadolinium

TRPC5icon pl   Patchliner data and applications: 
Data are taken from Milligan C.J. et al., Nature Protocols, 2009, 4(2), 244-255.

Current voltage relationships showing activaion of TRPC5 by extracellular application of Gd3+ (100 μM). Voltage ramps (−100 mV to 100 mV, holding potential 0 mV) were for 1s at 0.1 Hz.

KV7.5 - Current Voltage Recordings

PL KCNQ1icon pl   Patchliner data and applications:

Screenshot of KV7.5 recordings obtained on an eight-channel Patchliner. Currents are responses to an current voltage relationship type step pulse protocol.

 

 

 

 

Potassium Currents - Measuring Human Lymphoblasts

Lymphoblasticon pl   Patchliner data and applications:
Data are taken from Milligan C.J. et al., Nature Protocols, 2009, 4(2), 244-255.

An illustrative typical family of traces obtained during construction of a current voltage relationship from a human lymphoblast recording using amphotericin B (perforated patch). Currents were evoked by voltage steps from holding (−80 mV) to potentials varying from −100 mV to +60 mV.

TRPC1 (Smooth Muscle Cells) - Pharmacology

PL TRP SMCicon pl   Patchliner data and applications: 
Data are taken from Li, J. et al., Circulation Research, 2008,103(8), e97-104.

(a) Example whole cell recording showing current evoked by extracellular application of 2 μM thapsigargin (TG) and subsequent inhibition by 10 μM lanthanum (La3+). (b) As for (a) but showing the effect of 20 μg/ml anti-STIM1 antibody. (c) For the experiment shown in (b), curren voltage relationships for the current evoked by TG and blocked by anti-STIM1 antibody. (d) Mean currents normalized to pre-antibody values showing the effect of 20 μg/ml ani-STIM1 antibody and lack of effects for IgG or denatured anti-STIM1 antibody.

Glycine Receptor (GlyRa1 & GlyRa3) - Dose Response Analysis

PLGlycineLynch

icon pl   Patchliner data and applications:

(A) Effects of indicated concentrations of 3 on α1 and α3 GlyR currents activated by EC20 glycine concentrations as indicated. Unfilled bars denote glycine applications and filled bars denote compound applications. (B & C) Average dose response curves of 3 at α1 and α3 GlyRs, respectively.
Data are taken from Balansa W. et al., Bioorg Med Chem. 2010 Apr 15;18(8):2912-9.

 

Astrocytes - Analysing Potassium Currents

astrocytesicon pl   Patchliner data and applications:
Data are taken from Milligan C.J. et al., Nature Protocols, 2009, 4(2), 244-255

Left: Comparison of K+ current voltage relationships for rat astrocytes on the Patchliner (closed circles, n = 19) and on a conventional setup (open squares, n = 10). Currents were measured as a response to a voltage step protocol.
Right: Normalized K+ current amplitudes in rat astrocytes. Internal solution was changed to the same solution (K+, open circles, n = 7) or to one where Cs+ was substitued for K+ (Cs+,closed circles, n = 7). Currents were measured as responses to voltage steps from −100 mV to +40 mV.

TRPM3 - Pharmacology

icon pl   Patchliner data and applications:PL TRPM3 
Data are taken from Naylor J. et al., British Journal of Pharmacology, 2008, 1-7.

Shown are whole cell currents as responses to 0.2 s voltage ramps (−100 mV to +100 mV) which were applied every 10 s. All cells were HEK293 cells induced to express TRPM3. (a and c) Mean currents sampled at −80 and +80 mV, each normalized to the amplitude immediately before bath application of antiserum (TM3E3; 1:500 dilution) without (a, n = 8) or with (c, n = 4) pre-adsorption to 10 μM antigenic peptide. Pregnenolone sulphate (PregS) was bath applied at 25 μM. (b and d) Typical current voltage relationships from the experiments underlying (a) and (c).

KCa1.1 (BK) - Activation by Internal Calcium

BK verticalicon pl   Patchliner data and applications: 

Top: BK (KCa1.1) current voltage relationships in a single cell showing effects of changing the intracellular free Ca2+ concentration (15 nM, n = 9; 108 nM, n = 11; 316 nM, n = 11).

Bottom: Comparison of BK (KCa1.1) current voltage relationships obtained on a conventional patch clamp setup (closed circles, n = 10) and on the Patchliner (open circles, n = 11).
Data are taken from Milligan C.J. et al., Nature Protocols, 2009, 4(2), 244-255.

   

 

Glycine Receptor - Potentiation

application16 glycine 2

icon pl   Patchliner data and applications:
Cells and the positive modulator were kindly provided by Astrazeneca, Södertälje, Sweden.

Original traces of one application of 20 μM glycine followed by 6 applications of 20 μM glycine in conjunction with increasing concentrations of a positive modulator. 1 mM glycine was used as a second positive control.

 

Glycine Receptor (GlyRa1) - Antagonist

application16 glycine 3

icon pl   Patchliner data and applications:
Cells were kindly provided by Astrazeneca, Södertälje, Sweden.

The figure shows current responses of a hGlyRα1 expressing L-tk cell to alternating exposures to 100 μM glycine and 100 μM glycine + 1 μM strychnine.

 

GABAA Receptor (a1b2g2) - Activation

application16 gabaa 1 copy

icon pl   Patchliner data and applications:

Shown is concentration dependent activation of GABAA (α1β2γ2) receptors by 1, 3 and 10 μM GABA from a GABAA (α1β2γ2) expressing HEK293 cell. The rising phase in enlarged in the lower graph.

 

 

 

hERG - Pharmacology of Sotalol, Terfenadine, Verapamil (Results CiPA Phase I Study)

icon pl   CiPA PL phase 1 results 3Patchliner data and applications:
Cells were kindly provided by Charles River.

The effect of four compounds on hERG currents were investigated, using the CiPA voltage step protocol. Measured on the Patchliner the perforated patch methodology (Escin) and multi-hole chips (4 holes per well) were used. The IC50 value of Sotalol was determined as 157 µM, Terfenadine as 82.8 nM and Verapamil as 485 nM.

hERG - Pharmacology of Mexiletine, Quinidine, Ondansetron, Ranolazine (Results CiPA Phase I Study)

icon pl   CiPA PL phase 1 results 2Patchliner data and applications:
Cells were kindly provided by Charles River.

The effect of four compounds on hERG currents were investigated, using the CiPA voltage step protocol. Measured on the Patchliner the perforated patch methodology (Escin) and multi-hole chips (4 holes per well) were used. The IC50 value of Mexiletine was determined as 77.3 µM, Quinidine as 1.04 µM, Ondansetron as 1.13 µM and Ranolazine as 11.9 µM.

hERG - Pharmacology of Bepridil, Dofetilide, Cisapride, Diltiazem (Results CiPA Phase I Study)

icon pl   CiPA PL phase 1 results 1Patchliner data and applications:
Cells were kindly provided by Charles River.

The effect of four compounds on hERG currents were investigated, using the CiPA voltage step protocol. Measured on the Patchliner the perforated patch methodology (Escin) and multi-hole chips (4 holes per well) were used. The IC50 value of Cisapride was determined as 112 nM, Bepridil as 178 nM, Dofetilide as 33.9 nM and Diltiazem as 14.5 µM.

用户评价与案例分析

Dr. Carol Milligan - Statement about the Patchliner

icon pl   "In 2006 we purchased two of Nanion's very first Patchliner systems for our laboratories at the University of Leeds. We have been working with both systems very successfully for nearly four years now which is reflected in our recent high quality publications.
We operate both Patchliner systems on a daily basis, serving five different laboratories within the Faculty of Biological Sciences. Due to the diversity between the five groups, our Patchliners have been thoroughly put through their paces. This means that we have tested a large variety of different cells and a vast array of channel types, both endogenously and exogenously expressed. For example, we have investigated endogenous transient receptor potential (TRP) channels in human vascular smooth muscle cells and human synoviocytes. In addition, we have studied endogenous ion channels in primary astrocytes, human lymphoblasts and neutrophils, to mention a few.

Company Customers Carol 330The high data output has resulted in the production of an enormous amount of extremely good quality data using both native and primary mammalian cells. Our ability to make successful recordings from primary cells using the Patchliner is an extraordinary accomplishment, especially because the primary cell types that we work with are generally very difficult to record from using conventional patch clamp. Another impressive feature of this platform is its ability to perform routine intracellular perfusion which has enabled studies which might not have previously been attempted. We have also included our new technology as part of our annual 'Leeds Ion Channel Workshop' where it is well received. The Patchliner has turned out to be a show case project for us and has opened up new opportunities which I believe will play important roles in the future successes of research projects here in Leeds."

Dr. Carol Milligan, Research Fellow
Faculty of Biological Science, Leeds University, Leeds, UK

Dr. Chris Fanger - Statement about the Patchliner

icon pl   "We acquired the Patchliner because of its high data quality output combined with versatile and unique experimental features such as temperature control and internal solution exchange. In our TRP-channel initiative, we record from diverse ion channels and cells, and Patchliner quickly delivered accurate data with minimal assay development required. The data throughput is substantially increased by Patchliner’s innovative hardware and software, supporting short cycle times between experiments, minimal redundancy in compound screening and unlimited user control of the experiments. We are convinced that the Patchliner allows us to progress faster towards new discoveries, and at the same time it gives us a competitive edge because of its vast experimental flexibility".

Dr. Chris Fanger, Director of Lead Discovery
Hydra Biosciences, Boston, MA, USA

Dr. Thomas Seeger - Statement about the Patchliner

icon pl   “The Patchliner is a very good choice for the Bundeswehr research center. This instrument possesses features and offers great versatility that allow a broad  range of experimental protocols on diverse cell lines and ion channel targets. The Patchliner cooling plate allows not only cooling of cells to ensure high quality recordings over a couple of hours with the same cell batch, but it also allows to accurately store temperature sensitive supplements or test compounds over the complete experimental time during the day. The Patchliner is a great system for our many electrophysiology projects which are used to find new therapeutical options.”

Dr. Thomas Seeger
Bundeswehr Institute of Pharmacology and Toxicology, Munich, Germany

Marc Rogers - Statement about the Patchliner and CardioExcyte 96

icon pl   "It has been an absolute pleasure and privilege to work with Niels and the Nanion team over the past decade. I firmly believe that our two small, but independently minded ion channel focused companies, share a similar business ethos and a clear focus on delivering cost-effective solutions to our customers. In our extensive use of the Patchliner platform since 2007, my teams have benefited from the dedication of Nanion’s service engineers, application scientists, and bespoke data acquisition routines, whilst Nanion have benefited from seeing their products applied and adapted to real-world drug discovery projects and assays."

Icon CE   "This fruitful working relationship has now been extended to our use of the CardioExcyte96 phenotypic assay platform, which we utilise to develop and validate human iPSC cardiomyocyte cell lines and assays to meet the post-hERG, predictive cardiac safety testing needs of the drug discovery industry”."

Dr. Marc Rogers, CSO
Metrion Biosciences

Prof. Dr. David Weaver - Statement about the Patchliner

icon pl   “To invest in the Patchliner was a straightforward choice for the Vanderbilt Screening Center. This instrument combines a set of features that was particularly important to us. (...) Unlike other planar patch clamp devices we considered, the Patchliner gave us full access to the  electrophysiology modes we require for a broad range of experimental protocols including voltage clamp, current clamp, standard whole cell, cell attached, and perforated patch configurations combined with a facile ability to exchange the internal solution. (...) In a short time the Patchliner has already begun to expand Vanderbilt investigators’ horizons regarding what they can accomplish with electrophysiology.”

Dr. David Weaver, Research Associate Professor of Pharmacology, Director of the Chemical Biology's High-Throughput Screening Facility
Vanderbilt University, Nashville, TN, USA

Prof. Dr. Jerod Denton - Statement about the Patchliner

icon pl   "One of the biggest advantages of the Patchliner for our lab is that it enables students and fellows without formal training in patch clamp electrophysiology to begin generating meaningful data almost immediately. Instead of spending a couple of frustrating months learning the motor skills and hand-eye coordination necessary for conventional patch clamping with a microscope and micromanipulator, they spend that time designing and executing experiments, generating, evaluating and interpreting data and moving the project forward. The Patchliner also enables experimental flexibility that would be very difficult or impossible with conventional patch clamp rigs. For example, a talented undergraduate in the lab just finished up a series of intracellular drug application experiments. These studies would be difficult for the seasoned electrophysiologist and nearly impossible for an undergraduate student. However, the Patchliner allowed them to be completed in a very short period of time.

Company Customers Denton 330Another major advantage is that the Patchliner enables us to rapidly confirm or exclude hits from our primary fluorescence-based high-throughput screen using ­gold-standard electrophysiological methods. And the quality of the recordings rivals that of a conventional patch clamp rig. In fact, our first observations of pore "knock-off" of a new ROMK inhibitory small-molecule came from the Patchliner. These early observations from high-quality Patchliner recordings provided a clear ­direc­tion for subsequent mutagenesis work aimed at defining the binding site of this molecule within the cytoplasmic channel pore."

Jerod S. Denton, Ph.D.,Assistant Professor of Anesthesiology and Pharmacology
Vanderbilt University Medical Center, Nashville, TN, USA

网络研讨会与影像

12.09.2018 | 在线研讨会: CiPA study: Bridging ion channel and myocyte data


180912 event image CiPAII Webinar

Icon CE   CardioExcyte 96, icon pl   Patchliner and   icon sp96   SyncroPatch 384PE

日期: 9月12日, 4:00 PM CEST (北京时间10:00 PM)

 获取CiPA肌细胞与离子通道工作组的最新进展:

  • CiPA 肌细胞第二阶段验证研究结果:: cross-site comparison using the CardioExcyte 96
  • 高通量第一阶段研究: an update on progress of the CiPA Ion Channel Work Stream using the SyncroPatch 384PE and Patchliner

27.06.2017 | Webinar: New Dynamics in Automated Patch Clamp

icon pl   Patchliner

This webinar shows new applications on dynamic patch clamp of iPSC-derived cardiomyocytes and introduces an assay on KCa3.1 expressed in erythrocytes

28.07.2015 | Webinar: High Throughput and High Fidelity: Automated Patch Clamp in Screening and Research

icon sp96   SyncroPatch 384PE and   icon pl   Patchliner 

The webinar covers the use of the Patchliner and the SyncroPatch 384/768PE for characterization of ion channels and screening of ion channel active compounds.

11.05.2013 | Webinar: Patchliner - unlimited experimental freedom

icon pl   Patchliner   Patchliner - unlimited experimental freedom

This Webinar covers the features of the Patchlliner, including the "minimized cell usage" for expensive cells.

下载:

应用数据

Acetylcholine Receptor Alpha 3 Beta 4 - " Nicotinic a3b4 receptors recorded on Nanion's Patchliner"

icon pl   Patchliner application note:   logo pdf   (0.5 MB)

Acetylcholine Receptor Alpha 7 - "Human a7 nicotinic Acetylcholine Receptor on the Patchliner "

icon pl   Patchliner application note:   logo pdf   (0.2 MB)
Cells were kindly provided by Galantos.

AMPA receptor (GluA2) - "Activation, potentiation and inhibition of AMPA receptors on the Patchliner"

icon pl   Patchliner application note   logo pdf   (1.2 MB)

ASIC3 - "Characterization of hASIC3 (HEK) on Nanion´s Patchliner"

icon pl   Patchliner application note:   logo pdf   (0.6 MB)
Cells were kindly provided by Millipore.

Cardiac Ion Channels - "High Throughput Screening of Cardiac Ion Channels"

icon sp96   SyncroPatch 384PE   icon pl   Patchliner   Icon CE   CardioExcyte 96 application note   logo pdf   (2.3 MB)

Cardiomyocytes - "Combining automated patch clamp, impedance and EFP of hiPSC-CMs"

Icon CE   CardioExcyte 96   icon sp96   SyncroPatch 3984PE   icon pl   Patchliner Application Note 
Cells kindly provided by Takara-Clonetech.

Cardiomyocytes - "Recordings of action potentials in mouse ES cell-derived Cor.At cardiomyocytes on Nanion's Patchliner"

icon pl   Patchliner application note:   logo pdf   (0.7 MB)
Cells were kindly provided by Axiogenesis.  

Cardiomyocytes - "Voltage and current clamp recordings of Cellartis hiPS-CM on Nanion’s Patchliner"

icon pl   Patchliner application note:   logo pdf   (0.5 MB)
Cells were kindly provided by Takara Bio Europe AB.  

Cardiomyocytes - "Voltage and current clamp recordings of Cor.4U human iPS cell-derived cardiomyocytes on Nanion’s Patchliner"

icon pl   Patchliner application note:   logo pdf   (0.6 MB)
Cells were kindly provided by Axiogenesis.

CaV2.2 - "Characterization of CaV2.2 (HEK293) on Nanion's Patchliner"

icon pl   Patchliner application note:   logo pdf   (0.5 MB)

CaV3.2 - "Characterization of CaV3.2 (HEK293) on Nanion's Patchliner"

icon pl   Patchliner application note:   logo pdf   (0.6 MB)
Cells were kindly provided by Millipore.

CFTR - "Effect of internal F- on activation of Cystic Fibrosis Transmembrane Conductance (CFTR) regulator by forskolin " (report)

icon pl   Patchliner application report:   logo pdf   (0.2 MB)

CFTR - "Effect of internal F- on activation of Cystic Fibrosis Transmembrane Conductance (CFTR) regulator by forskolin"

icon pl   Patchliner application note:   logo pdf   (0.7 MB)

GABAA (a1b2g2) - "Patchliner - Investigating the pharmacology of GABAA receptors expressed in HEK293 cells"

icon pl   Patchliner application note:   logo pdf   (0.7 MB)

GABAA (a1b2g2) - "Rapid agonist applications in precise intervals (GABAA) "

icon pl   Patchliner application note:   logo pdf   (1.5 MB)

Glycine receptors (GlyRa1) - "Modulators for glycine receptors investigated using the Patchliner "

icon pl   Patchliner application note:   logo pdf   (0.4 MB)
Cells were kindly provided by AstraZeneca.

hERG - "Effect of temperature on erythromycin action on hERG currents recorded on Nanion's Patchliner"

icon pl   Patchliner application note:   logo pdf   (0.9 MB)
Cells were kindly provided by Millipore.

KCa3.1 - "Modulation of hKCa3.1 by internal Ca2+ performed on Nanion’s Patchliner"

icon pl   Patchliner application note:   logo pdf   (0.6 MB)
Cells were kindly provided by Charles River.

KV1.3 - "Pharmacological analysis of heterologous and endogenous expressed KV1.3 channels in Sf21 insect cells and T-lymphocytes"

icon pl   Patchliner application note:   logo pdf   (0.7 MB)
Cells were kindly provided by conoGenetix.

NaV1.5 - "Characterization of CreaCell's hNaV1.5 (A-0822) on Nanion's Patchliner"

icon pl   Patchliner application note:   logo pdf   (0.9 MB)
Cells were kindly provided by CreaCell.

NaV1.5 - "Pharmacology of hNaV1.5 recorded on Nanion's Patchliner"

icon pl   Patchliner application note:   logo pdf   (0.3 MB)
Cells were kindly provided by Millipore.

NaV1.7 - "Pharmacology on hNaV1.7 performed on Nanion’s Patchliner at Vhalf "

icon pl   Patchliner application note:   logo pdf   (0.4 MB)
Cells were kindly provided by Anaxon.

NaV1.8 - "Characterization of rNaV1.8 (ND7-23) on Nanion's Patchliner"

icon pl   Patchliner application note:   logo pdf   (0.4 MB)
Cells were kindly provided by Millipore.

Neurons - "Peri.4U and Dopa.4U stem cell-derived neurons recorded on Nanion´s Patchliner"

icon pl   Patchliner application note:   logo pdf   (0.6 MB)
Cells were kindly provided by Axiogenesis.  

Neurons - "Stem cell-derived neurons recorded on Nanion's Patchliner"

icon pl   Patchliner application note:   logo pdf   (0.4 MB)
Cells were kindly provided by Cellular Dynamics.

P2X2 / P2X3 - "P2X2/3 receptors recorded on Nanion's Patchliner"

icon pl   Patchliner application note:   logo pdf   (0.4 MB)
Cells were kindly provided by Evotec.  

TRPM8 - "TRPM8 activation by menthol and Eucalyptol performed on Nanion’s Patchliner"

icon pl   Patchliner application note:   logo pdf   (0.4 MB)
Cells were kindly provided by Charles River.

TRPV1 - "Heat activation of TRPV1 on Nanion's Patchliner"

icon pl   Patchliner application note:   logo pdf   (0.5 MB)

TRPV3 - "Heat activation of TRPV3 on Nanion's Patchliner"

icon pl   Patchliner application note:   logo pdf   (0.6 MB)

产品彩页

Patchliner - 产品单页

icon pl   Patchliner 产品单页:    logo pdf   (0.8 MB)

Patchliner 产品彩页 - Cooling Plate

icon pl   Patchliner 产品彩页:    logo pdf   (3.3 MB)

Patchliner 产品彩页 - PatchControl HT Release 2017

icon pl   Patchliner 产品彩页:    logo pdf   (4.6 MB)

发表文献

2018 - Selective NaV1.1 activation rescues Dravet syndrome mice from seizures and premature death

icon pl   Patchliner publication in PNAS (2018)

Authors:
Richards K.L., Milligan C.J., Richardson R.J., Jancovski N., Grunnet M., Jacobson L.H., Undheim E.A.B., Mobli M., Chow C.Y., Herzig V., Csoti A., Panyi G., Reid A.A., King G.F., Petrou S.

2018 - Electrophysiological investigation of the effect of structurally different bispyridinium non-oxime compounds on human α7-nicotinic acetylcholine receptor activity - An in vitro structure-activity analysis

icon pl   Patchliner publication in Toxicology Letters (2018)

Authors:
Scheffel C., Niessen K.V., Rappenglück S., Wanner K.T., Thiermann H., Worek F., Seeger T.

2018 - Discovery of a small molecule modulator of the Kv1.1/Kvβ1 channel complex that reduces neuronal excitability and in vitro epileptiform activity

icon pl   Patchliner publication in CNS Neuroscience & Therapeutics (2018)

Authors:
Niespodziany I., Mullier B., André V.M., Ghisdal P., Jnoff E., Moreno‐Delgado D., Swinnen D., Sands Z., Wood M., Wolff C.

2018 - Dehydroevodiamine and hortiamine, alkaloids from the traditional Chinese herbal drug Evodia rutaecarpa, are IKr blockers with proarrhythmic effects in vitro and in vivo

icon pl   Patchliner publication in Pharmacological Research (2018)

Authors:
Baburin I., Varkevisser R., Schramm A., Saxena P., Beyl S., Szkokan P., Linder T., Stary-Weinzinger A., van der Heyden M.A.G., Houtman M., Takanari H., Jonsson M., Beekman J.H.D., Hamburger M., Vos M.A., Hering S.

2018 - Counteracting desensitization of human α7-nicotinic acetylcholine receptors with bispyridinium compounds as an approach against organophosphorus poisoning

icon pl   Patchliner publication in Toxicology Letters (2018)

Authors:
Scheffel C., Niessen K.V., Rappenglück S., Wanner K.T., Thiermann H., Worek F., Seeger T.

2018 - An update on the advancing high-throughput screening techniques for patch clamp-based ion channel screens: implications for drug discovery

icon sp96   SyncroPatch 384/768PE and   icon pl   Patchliner publication in Expert Opinion on Drug Discovery

Authors: 
Obergrussberger A., Goetze T.A., Brinkwirth N., Becker N., Friis S., Rapedius M., Haarmann C., Rinke-Weiß I., Stölzle-Feix S., Brüggemann A., George M., Fertig N.

2018 - A Previously Unrecognized Ca2+-inhibited Nonselective Cation Channel in Red Blood Cells

icon pl   Patchliner publication in HemaSphere (2018)

Authors:
Petkova-Kirova P., Hertz L., Makhro A., Danielczok J., Huisjes R., Llaudet-Planas E., Mañú-Pereira, M., Vives Corrons J.-L., van Wijk R., Bogdanova A., Kaestner L.

2018 - A Hybrid Model for Safety Pharmacology on an Automated Patch Clamp Platform: Using Dynamic Clamp to Join iPSC-Derived Cardiomyocytes and Simulations of IK1 Ion Channels in Real-Time

icon pl  Patchliner publication in Frontiers in Physiology

Authors: 
Goversen B., Becker., N., Stölzle-Feix S., Obergrussberger A., Vos M.A., van Veen T.A.B., Fertig N., de Boer T.P.

2017 - Trifluoperazine-Induced Suicidal Erythrocyte Death and S-Nitrosylation Inhibition, Reversed by the Nitric Oxide Donor Sodium Nitroprusside

icon pl   Patchliner publication in Cellular Physiology and Biochemistry (2017)

Authors:
Ghashghaeinia M., Wesseling M.C., Ramos E., Petkova-Kirova P., Waibel S., Lang E., Bissinger R., Alzoubi K., Edelmann B., Hosseinzadeh Z., Dreischer P., Shahvaroughi-Farahani A., Mrowietz U., Köberle M., Kaestner L., Bernhardt I., Martínez-Ruiz A., Wieder T., Lang F.

2017 - Purinergic receptors P2RX4 and P2RX7 in familial multiple sclerosis

icon pl  Patchliner publication in Human Mutation (2017)

Authors: 
Sadovnick A.D., Gu B.J., Traboulsee A.L., Bernales C.Q., Encarnacion M., Yee I.M., Criscuoli M.G., Huang X., Ou A., Milligan C.J., Petrou S, Wiley J.S, Vilariño-Güell C.

2017 - Myoclonus Epilepsy and Ataxia due to KCNC1 Mutation: Analysis of 20 Cases and K+ Channel Properties

icon pl  Patchliner publication in Annals in Neurobiology (2017)

Authors: 
Oliver K.L., Franceschetti S., Milligan C.J., Muona M., Mandelstam S.A., Canafoglia L., Boguszewska-Chachulska A.M., Korczyn A.D.,Bisulli F., Di Bonaventura C., Ragona F., Michelucci R., Ben-Zeev B., Straussberg R., Panzica F., Massano J., Friedman D., Crespel A., Engelsen B.A., Andermann F., Andermann E., Spodar K., Lasek-Bal A., Riguzzi P.,Pasini E., Tinuper P., Licchetta L., Gardella E., Lindenau M., Wulf A., Møller R.S.

2017 - L-Type Calcium Channel Inhibition Contributes to the Proarrhythmic Effects of Aconitine in Human Cardiomyocytes

icon pl  Patchliner publication in PLoS ONE (2017)

Authors: 
Wu J., Wang X., Chung Y.Y. Koh C.H. Liu Z., Guo H., Yuan Q., Wang C., Su S., Wei H.

2017 - Discovery of benzimidazole derivatives as modulators of mitochondrial function: A potential treatment for Alzheimer's disease

icon pl  Patchliner publication in PLoS ONE (2017)

Authors: 
Kim T., Yang H.Y., Park B.G., Jung S.Y., Park J.H., Park K.D., Min S.J., Tae J., Yang H., Cho S., Cho S.J., Song H., Mook-Jung I., Lee J., Pae A.N.

2017 - Development of Automated Patch Clamp Technique to Investigate CFTR Chloride Channel Function

icon pl  Patchliner publication in Frontiers in Pharmacology (2017)

Authors: 
Billet A.,Froux L., Hanrahan J.W., Becq F.

2017 - Correlation between human ether-a-go-go related gene channel inhibition and action potential prolongation

icon pl   Patchliner publication in British Journal of Pharmacology (2017)

Authors:
Saxena P., Hortigon‐Vinagre M.P., Beyl S.,Baburin I., Andranovits S., Iqbal S.M., Costa A., IJzerman A.P., Kügler P., Timin E., Smith G.L., Hering S.

2017 - Automated Patch Clamp Recordings of Human Stem Cell- Derived Cardiomyocytes.

icon pl  Patchliner and   icon sp96   SyncroPatch 384PE book chapter in Stem Cell-Derived Models in Toxicology (2017)

Authors: 
Obergrussberger A., Haarmann C., Stölzle-Feix S., Becker N., OhtsukiA., Brüggemann A., George M., Fertig N.

2017 - 14-3-3 proteins regulate K2P5.1 surface expression on T lymphocytes

icon pl  Patchliner publication in Traffic (2017)

Authors:
Fernández-Orth J., Ehling P., Ruck T., Pankratz S., Hofmann M.,Landgraf P., Dieterich D.C., SmallaK.-H., Kähne T., Seebohm G., Budde T., Wiendl H., Bittner S., Meuth S.G.

2017 - 'Gardos Channelopathy': a variant of hereditary Stomatocytosis with complex molecular regulation

icon pl  Patchliner publication in Scientific Reports (2017)

Authors: 
Fermo E., Bogdanova A., etkova-Kirova P., Zaninoni A., Marcello A.P., Makhro A., Hänggi P., Hertz L., Danielczok J., Vercellati C., Mirra N., Zanella A., Cortelezzi A., Barcellini W., Kaestner L., Bianchi P.

2016 - The antiepileptic medications carbamazepine and phenytoin inhibit native sodium currents in murine osteoblasts

icon pl  Patchliner publication in Epilepsia (2016)

Authors: 
Petty S.J., Milligan C.J., Todaro M., Richards K.L., Kularathna P.K., Pagel C.N., French C.R., Hill-Yardin E.L., O'Brien T.J., Wark J.D., Mackie E.J., Petrou S.

2016 - Myrsinane, Premyrsinane, and Cyclomyrsinane Diterpenes fromEuphorbia falcata as Potassium Ion Channel Inhibitors with Selective G Protein-Activated Inwardly Rectifying Ion Channel (GIRK) Blocking Effects

icon pl  Patchliner publication in Journal of Natural Products (2016)

Authors: 
Vasas A., Forgo P., Orvos P., Tálosi L., Csorba A., Pinke G., Hohmann J.

2016 - Human T cells in silico: Modelling their electrophysiological behaviour in health and disease

icon pl  Patchliner publication in Journal of Theoretical Biology (2016)

Authors: 
Ehling P., Meuth P., Eichinger P., Hermann A.M., Bittner S., Pawlowski M., Pankratz S., Herty M., Budde T., Meuth S.G.

2016 - Human EAG channels are directly modulated by PIP2 as revealed by electrophysiological and optical interference investigations

icon pl  Patchliner publication in Scientific Reports (2016)

Authors: 
Han B., He K., Cai C., Tang Y., Yang L., Heinemann S.H., Hoshi T., Hou S.

2016 - Coupling Data Mining and Laboratory Experiments to Discover Drug Interactions Causing QT Prolongation

icon pl  Patchliner publication in Journal of the American College of Cardiology (2016)

Authors:
Lorberbaum T., Sampson K.J., Chang J.B., Iyer V., Woosley R.L., Kass R.S., Tatonetti N.P.

2016 - Automated Electrophysiological and Pharmacological Evaluation of Human Pluripotent Stem Cell-Derived Cardiomyocytes

icon pl  Patchliner publication in Stem Cells and Development (2016)

Authors: 
Rajamohan D., Kalra S., Hoang M.D., George V., Staniforth A., Russell H., Yang X., Denning C.

2015 - Scalable Electrophysiological Investigation of iPS Cell-Derived Cardiomyocytes Obtained by a Lentiviral Purification Strategy

icon pl  Patchliner publication in Journal of Clinical Medicine (2015)

Authors: 
Friedrichs S., Malan D., Voss Y., Sasse P.

2015 - Novel screening techniques for ion channel targeting drugs

icon pl  Patchliner,   icon sp96   SyncroPatch 384PE and   Icon CE   CardioExcyte 96 publication in Channels (2015)

Authors: 
Obergrussberger A., Stölzle-Feix S., Becker N., Brüggemann A., Fertig N., Möller C.

2014 - Physiological and genetic analysis of multiple sodium channel variants in a model of genetic absence epilepsy

icon pl  Patchliner publication in Neurobiology of Disease (2014)

Authors: 
Oliva M.K., McGarr T.C., Beyer B.J., Gazina E., Kaplan D.I., Cordeiro L., Thomas E., Dib-Hajj S.D., Waxman S.G., Frankel W.N., Petrou S.

2014 - Multi-Generational Pharmacophore Modeling for Ligands to the Cholane Steroid-Recognition Site in the β1 Modulatory Subunit of the BK(Ca) Channel

icon pl  Patchliner publication in Journal of Molecular Graphics and Modelling (2014)

Authors: 
McMillan J.E., Bukiya A.N., Terrell C.L., Patil S.A., Miller D.D., Dopico A.M., Parrilla A.L.

2014 - Early identification of hERG liability in drug discovery programs by automated patch clamp

icon pl  Patchliner and   icon sp96   SyncroPatch 384PE publication in Frontiers in Pharmacology (2014)

Authors: 
Danker T., Moeller C.

2014 - Automated Patch Clamp Analysis of nAChα7 and NaV1.7 Channels

icon pap  Port-a-Patch and   icon pl   Patchliner publication in Current Protocols in Pharmacology (2014)

Authors: 
Obergrussberger A., Haarmann C., Rinke I., Becker N., Guinot D., Brueggemann A., Stoelzle-Feix S., George M., Fertig N.

2013 - Red cell investigations: Art and artefacts

icon pl  Patchliner publication in Blood reviews (2013)

Authors: 
Minetti G., Egée S., Mörsdorf D., Steffen P., Makhro A., Achilli C., Ciana A., Wang J., Bouyer G., Bernhardt I., Wagner C., Thomas S., Bogdanova A., Kaestner L.

2013 - Minimized cell usage for stem cell-derived and primary cells on an automated patchclamp system

icon pl  Patchliner publication in Journal of Pharmacological and Toxicological Methods (2013)

Authors: 
Becker N., Stoelzle S., Göpel S., Guinot D., Mumm P., Haarmann C., Malan D., Bohlen H., Kossolov E., Kettenhofen R., George M., Fertig N., Brüggemann A.

2013 - Establishment of a Secondary Screening Assay for P/Q-Type Calcium Channel Blockers

icon pl   Patchliner publication in Combinatorial Chemistry & High Throughput Screening (2013)

Authors: 
Hermann D., Mezler M., Swensen A., Bruehl C., Obergrußberger A., Wicke K., Schoemaker H., Gross G., Draguhn A., Nimmrich V.

2013 - Automated Planar Patch Clamp

icon pl   Patchliner book chapter in Ion Channels (2013)

Authors: 
Milligan, C. J., Möller, C.

2012 - TRPA1 Agonist Activity of Probenecid Desensitizes Channel Responses: Consequences for Screening

icon pl   Patchliner publication in ASSAY and Drug Development Technologies (2012)

Authors: 
McClenaghan C., Zeng F., Verkuyl J.M.

2012 - Toward a new gold standard for early safety: automated temperature-controlled hERG test on the Patchliner

icon pl   Patchliner publication in Frontiers in Pharmacology (2012)

Authors: 
Polonchuk L.

2012 - Synthesis and In Vitro Antibacterial Activity of Novel 3‐Azabicyclo [3.3. 0] octanyl Oxazolidinones

icon pl   Patchliner publication in Chemical Biology and Drug Design (2012)

Authors: 
Bhattarai D., Lee S.H., Seo S.H., Nam G., Kang S.B., Pae A.N., Kim E.E., Oh T., Cho S.N., Keum G.

2012 - Observation of Antinociceptive Effects of Oxymatrine and its Effect on Delayed Rectifier K+ Currents (Ik) in PC12 Cells

icon pl   Patchliner publication in Neurochemical Research (2012)

Authors: 
Wang Y., Yuan J., Yuan X., Wang W., Pei X., Zhao Q., Cao H., Xu M., Liu Z.

2012 - Natural and artificial ion channels for biosensing platforms

icon pap   Port-a-Patch,   icon pl   Patchliner,   icon sp96   SyncroPatch 96 ((a predecessor model of SyncroPatch 384PE) and   icon vpp   Vesicle Prep Pro publication in Analytical and Bioanalytical Chemistry (2012)

Authors: 
Steller L., Kreir M., Salzer R.

2012 - HTS techniques for patch clamp-based ion channel screening - economy and advances

icon pap   Port-a-Patch,   icon pl   Patchliner and   icon sp96   SyncroPatch 96 (a predecessor model of SyncroPatch 384PE) publication in Expert Opinion on Drug Discovery (2012)

Authors: 
Farre C. and Fertig N.

2012 - Characterizing Human Ion Channels in Induced Pluripotent Stem Cell-Derived Neurons

icon pl   Patchliner publication in Journal of Biomolecular Screening (2012)

Authors: 
Haythornthwaite A, Stoelzle S, Hasler A, Kiss A, Mosbacher J, George M, Brüggemann A, Fertig N.

2011 - State-of-the-art automated patch clamp devices: heat activation, action potentials, and high throughput in ion channel screening

icon pap   Port-a-Patch,   icon pl  Patchliner and   icon sp96   SyncroPatch 96 (a predecessor model of SyncroPatch 384PE) publication in Frontiers in Pharmacology (2011)

Authors: 
Stoelzle S., Obergrussberger A., Brüggemann A., Haarmann C., George M., Kettenhofen R., Fertig N.

2011 - Rapid and Contrasting Effects of Rosiglitazone on Transient Receptor Potential TRPM3 and TRPC5 Channels

icon pl  Patchliner publication in Molecular Pharmacology (2011)

Authors: 
Majeed Y., Bahnasi Y., Seymour V.A.L., Wilson L.A., Milligan C.J., Agarwal A.K., Sukumar P., Naylor J., Beech D.J.

2011 - Pharmacological studies of Cav3.1 T-type calcium channels using automated patch-clamp techniques

icon pl  Patchliner publication in General Physiology and Biophysics (2011)

Authors: 
Choi K.-H., Song C., Cheong C.S., Rhim H.

2011 - Development of a selective small-molecule inhibitor of Kir1.1, the Renal Outer Medullary Potassium Channel

icon pl  Patchliner publication in Molecular Pharmacology (2011)

Authors: 
Bhave G., Chauder B.A., Liu W., Dawson E.S., Kadakia R., Nguyen T.T., Lewis L.M., Meiler J., Weaver C.D., Satlin L.M., Lindsley C.W., Denton J.S.

2011 - Cardiac Toxicity

icon pl   Patchliner and   icon pap   Port-a-Patch book chapter in ADMET for Medicinal Chemists: A Practical Guide (2011)

Authors: 
Kettenhofen R., Schwengberg S.

2011 - Automated Patch Clamp on mESC-Derived Cardiomyocytes for Cardiotoxicity Prediction

icon pl   Patchliner and   icon pap   Port-a-Patch publication in Journal of Biological Chemistry (2011)

Authors: 
Stoelzle S., Haythornthwaite A., Kettenhofen R., Kolossov E., Bohlen H., George M., Brüggemann A., Fertig N.

2011 - Automated electrophysiology makes the pace for cardiac ion channel safety screening

icon pl  Patchliner and   icon sp96   SyncroPatch 96 (a predecessor model of SyncroPatch 384PE) publication in Frontiers in Pharmacology (2011)

Authors: 
Möller C., Witchel H.

2011 - The Chimeric approach reveals that differences in the TRPV1 pore domain determine species-specific sensitivity to block of heat activation

icon pl   Patchliner publication in Journal of Biological Chemistry (2011)

Authors: 
Papakosta M., Dalle C., Haythornthwaite A., Cao L., Stevens E.B., Burgess G., Russell R., Cox P.J., Phillips S.C., Grimm C.

2010 - Renaissance of ion channel research and drug discovery by patch clamp automation

icon pap  Port-a-Patch,   icon pl   Patchliner and   icon sp96   SyncroPatch 96 (a predecessor model of SyncroPatch 384PE)  publication in Future Medical Chemistry (2010)

Authors:
Farre C. and Fertig N.

2010 - Propranolol blocks cardiac and neuronal voltage-gated sodium channels

icon pl  Patchliner publication in Frontiers in Pharmacology (2010)

Authors: 
Wang D.W., Mistry A.M., Kahlig K.M., Kearney J.A, Xiang J., George Jr. A.L.

2010 - Pregnenolone Sulphate- and Cholesterol-Regulated TRPM3 Channels Coupled to Vascular Smooth Muscle Secretion and Contraction

icon pl  Patchliner publication in Circulation Research (2010)

Authors: 
Naylor J., Li J., Milligan C.J, Zeng F., Sukumar P., Hou B., Sedo A., Yuldasheva N., Majeed Y., Beri D., Jiang S., Seymour V.A.L, McKeown L., Kumar B., Harteneck C., O'Regan D., Wheatcroft S.B., Kearney M.T, Jones C., Porter K.E., Beech D.J.

2010 - Multiplexed transposon-mediated stable gene transfer in human cells

icon pl  Patchliner publication in Proc. Natl. Acad. Sci. USA. (2010)

Authors: 
Kahlig K.M., Saridey S.K., Kaja A., Daniels M.A., George A.L. Jr., Wilson M.H.

2010 - Ircinialactams: subunit-selective glycine receptor modulators from Australian sponges of the family Irciniidae

icon pl  Patchliner publication in Bioorganic & Medical Chemistry (2010)

Authors: 
Balansa W, Islam R, Fontaine F, Piggott AM, Zhang H, Webb TI, Gilbert DF, Lynch JW, Capon RJ.

2010 - Cor.At Cardiomyocytes: Primary-like Cardiomyocytes for Manual and Automated Electrophysiological Screening

icon pap  Port-a-Patch and   icon pl   Patchliner publication in Lonza Resource Notes (2010)

Authors:
Kettenhofen R., Stölzle S.

2009 - Robotic multiwell planar patch-clamp for native and primary mammalian cells

icon pl  Patchliner publication in Nature Protocols (2009)

Authors:
Milligan C.J., Li J., Sukumar P., Majeed Y., Dallas M.L., English A., Emery P., Porter K.E., Smith A.M., McFadzean I., Beccano-Kelly D., Bahnasi Y., Cheong A., Naylor J., Zeng F., Liu X., Gamper N., Jiang L., Pearson H.A., Peers C., Robertson B., Beech D.J.

2009 - Port-a-Patch and Patchliner: High fidelity electrophysiology for secondary screening and safety pharmacology

icon pap  Port-a-Patch and   icon pl   Patchliner publication in Combinatorial Chemistry & High Throughput Screening (2009)

Authors: 
Farre C., Haythornthwaite A., Haarmann C., Stoelzle S., Kreir M., George M., Brüggemann A., Fertig N.

2009 - High-throughput screening reveals a small-molecule inhibitor of the renal outer medullary potassium channel and Kir7.1

icon pl  Patchliner publication in Molecular Pharmacology (2009)

Authors: 
Lewis L.M., Bhave G., Chauder B.A., Banerjee S., Lornsen K.A., Redha R., Fallen K., Lindsley C.W., Weaver C.D., Denton J.S.

2009 - High throughput techniques for discovering new glycine receptor modulators and their binding sites

icon pl  Patchliner publication in Frontiers in Molecular Neuroscience (2009)

Authors: 
Gilbert D.F.,Islam R., Lynagh T., Lynch J.W., Webb T.I. Front.

2008 - TRPC channel activation by extracellular thioredoxin

icon pl  Patchliner publication in Nature (2008)

Authors: 
Xu S.Z., Sukumar P., Zeng F., Li J., Jairaman A., English A., Naylor J., Ciurtin C., Majeed Y., Milligan C.J., Bahnasi Y.M., Al-Shawaf E., Porter K.E., Jiang L.H., Emery P., Sivaprasadarao A., Beech D.J.

2008 - Production of a specific extracellular inhibitor of TRPM3 channels

icon pl  Patchliner publication in British Journal of Pharmacology (2008)

Authors: 
Naylor J., Milligan C.J., Zeng F., Jones C., Beech D.J.

2008 - Ion channel screening – automated patch clamp on the rise

icon pap  Port-a-Patch and   icon pl   Patchliner publication in Drug Discovery Today (2008)

Authors: 
Farre C., George M., Brüggemann A., Fertig N.

2008 - Interactions, functions, and independence of plasma membrane STIM1 and TRPC1 in vascular smooth muscle cells

icon pl  Patchliner publication in Circulation Research (2008)

Authors: 
Li J., Sukumar P., Milligan C.J., Kumar B., Ma Z.Y., Munsch C.M., Jiang L.H., Porter K.E., Beech D.J.

2007 - Planar Patch Clamping

icon pap  Port-a-Patch and   icon pl   Patchliner book chapter in "Patch Clamp Analysis – Advanced Techniques", Series: Neuromethods (2007)

Authors: 
Behrends, J.C., Fertig, N.

2007 - Automated ion channel screening: patch clamping made easy

icon pap  Port-a-Patch and   icon pl   Patchliner publication in Expert Opinion Therapeutic Targets (2007)

Authors: 
Farre C., Stoelzle S., Haarman C., George M., Brueggemann A., Fertig N.

2006 - Microchip technology for automated and parallel patch clamp recording

icon pap  Port-a-Patch and   icon pl   Patchliner publication in Small Journal (2006)

Authors: 
Brüggemann A., Stoelzle S., George M., Behrends J.C., Fertig N.

海报

2018 - Expression and pharmacology of GluA2-containing AMPA receptors in cell lines and stem cell-derived neurons

icon pap   Port-a-Patch,   icon pl   Patchliner and   icon sp96   SyncroPatch 384PE poster, Europhysiology Meeting 2018  logo pdf   (0.9 MB)

2018 - Combining electrophysiology and contractility recordings for more complete assessment of hiPSC-CMs

icon sp96   SyncroPatch 384PE,   icon pl   Patchliner and   Icon CardioExcyte 96 simpel RGB   CardioExcyte 96 poster, Europhysiology Meeting 2018  logo pdf   (1.4 MB)

2017 - Cardiomyocytes in Voltage Clamp and Current Clamp by Automated Patch Clamp

icon sp96   SyncroPatch 384PE and   icon pl   Patchliner poster, BPS Meeting 2017  logo pdf   (1.7 MB)

2018 - Investigating pain pathways by inhibition of voltage-gated sodium channels

icon sp96   SyncroPatch 384PE and   icon pl   Patchliner poster, FENS Meeting 2018  logo pdf   (2.5 MB)

2018 - Introducing simulated IK1 into human iPSC-cardiomyocytes using dynamic clamp on an automated patch clamp setup

icon pl   Patchliner poster, Biophysics Annual Meeting 2018  logo pdf   (1.0 MB)

2015 - Complementary automated patch clamp, extracellular field potential and impedance recordings of iPSCs: safety screening tool box for the future

icon pl   Patchliner and   Icon CE   CardioExcyte 96 and   icon sp96   SyncroPatch 384PE poster,   SPS 2015   logo pdf   (2.7 MB)

2015 - The backstage pass to study your favorite TRP channel

icon pap   Port-a-Patch and   icon pl   Patchliner and SyncroPatch 384PE and   icon sp96   SyncroPatch 384PE poster, TRP Meeting 2015   logo pdf   (2.2 MB)

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