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CaV2.2 - Current-to-Voltage Relationship

Cav22IV PaP

icon pap   Port-a-Patch data and applications:
Cells were kindly provided by Millipore.

Representative current responses of an individual HEK-293 cell expressing CaV2.2 to a standard voltage protocol. Average current-voltage relationship (n = 10). The error bars reflect the standard error of the mean (S.E.M.).

2016 - Automated Patch Clamp Meets High-Throughput Screening: 384 Cells Recorded in Parallel on a Planar Patch Clamp Module

icon sp96  SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) publication in Journal of Lab Automation (2016)

Authors: 
Obergrussberger A., Brüggemann A., Goetze T.A., Rapedius M., Haarmann C., Rinke I., Becker N., Oka T., Ohtsuki A., Stengel T., Vogel M., Steindl J., Mueller M., Stiehler J., George M., Fertig N.

2015 - Electrophysiological analysis of mammalian cells expressing hERG using automated 384-well-patch-clamp

icon sp96  SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) publication in BCM Pharmacology and Toxicology (2015) 

Authors: 
Haraguchi Y., Ohtsuki A., Oka T., Shimizu T.

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.

GABAA Receptor (α1β2γ2) - Success Rates

Seal Stat GABAa1

icon sp96   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.

 

 

 

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).

 

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.

 

NaV1.5 - Screening Online Analysis

SyncroPatch NaOA small

icon sp96   SyncroPatch 96 (a predecessor model of SyncroPatch 384PE) data and applications:
Cells were kindly provided by Millipore.

The image shows the state dependent block at -130 mV (light blue) and -90 mV (dark blue) by the addition of increasing compound concentrations to the different recording wells. The large window shows the effect of increasing concentrations of Lidocaine (red box) on the hNav1.5 currents.

NaV1.5 - Screening Mode

SyncroPatch NaTrace small

icon sp96   SyncroPatch 96 (a predecessor model of SyncroPatch 384PE) data and applications:
Cells were kindly provided by Millipore.

Compound effect on hNav1.5  was investigated. A two-pulse protocol (-90 mV / -130 mV) was used to establish if the different compounds blocked the Nav1.5 currents in a state-dependent manner.
The image shows raw data traces. Success rate for the experiment was 71% (>1 GOhm seal resistance), indicated by the green color.
The corresponding online analysis of this experiment is shown in the data set NaV1.5 - Screening Online Analysis.

NaV1.5 - Inactivation Protocol

NaInactivation

icon sp96   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 - Lidocaine Dose Response

NaLidocaine

icon sp96   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 potentials (as indicated) either using a single concentration of compound 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.

CaV3.2 - Raw Currents

SyncroCav32Currents

icon sp96   SyncroPatch 96 (a predecessor model of SyncroPatch 384PE) data and applications:
Cells were kindly provided by Cytomyx Millipore, UK.

Shown are raw current responses of CaV3.2 (HEK293) to a current voltage relationship step protocol. 75 % of the cells had a seal above 0.5 GΩ (color coded in green). The corresponding online analysis of this experiment is shown in the data set CaV3.2 - Online Analysis.

 

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.

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. 

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!

hERG - Pharmacological Experiments at 35 °C

p28 4 fullDoseRespicon pap   Port-a-Patch data and applications:
Cells were kindly provided by Cytomyx/Millipore, UK.

A full dose response curve of quinidine acting on the hERG channel was obtained at physiological temperature (35 °C). The IC50 for quinidine at physiological temperature was 1.3 ± 0.2 μM (n = 5), similar to that obtained at room temperature (1.0 ± 0.03 μM, n = 3).

NaV1.5 - Accurate Voltage Control

p14 4 Nav

icon pap   Port-a-Patch data and applications:
Cells stably transfected with human SCN5A were kindly provided by Millipore.

To perform recordings of fast events, such as the activation and inactivation of Na currents, it is essential to have accurate voltage control. The image shows currents and I/V characteristics of NaV1.5 expressed in HEK293 cells.

CaV3.2 - Channel Blockers

Cav32 Block

icon pap   Port-a-Patch data and applications:
Cells were kindly provided by Cytomyx Millipore.

Shown are raw current traces (top) and average dose response curves (bottom) of CaV3.2 (T-type Ca2+- Channel) block by compounds as indicated. CaV3.2 is stably expressed in HEK293 cells. IC50s were 863 nM (Mibefradil), 27 μM (Nifedipine) and 52 μM Amiloride.

 

 

 

 

CaV3.2 - T-Type Calcium Channels

Cav32 IV

icon pap   Port-a-Patch data and applications:
Cells were kindly provided by Cytomyx Millipore.

Shown are raw current traces (top left) and average peak current data (top right) of the current voltage relationship of CaV3.2 (T-type Ca2+- Channel) stably expressed in HEK293 cells. Activation and Inactivation plots were constructed (bottom). Half-activation and half-inactivating potentials were determined as -32 mV and -65 mV, respectively.

 

 

hERG - Good Results with "Sticky Compounds"

p14 1 hERG

icon pap   Port-a-Patch data and applications:
Cells were kindly provided by Cytomyx/Millipore.

Sticky compounds, such as some hERG blockers, exhibit expected IC50 values with the Port-a-Patch. The concentrations of the half maximal block were: 1.27 nM (astemizole), 8.9 nM (cisapride), 163.7 nM (flunarizine) and 11.0 nM (terfenadine).

 

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

 

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.

ASIC3 - "Activation and Inhibition of human ASIC3 Channels on Nanion’s SyncroPatch 384PE"

icon sp96   SyncroPatch 384PE (a predecessor model of SyncroPatch 384) application note:   logo pdf   (2.2 MB)
Cells were kindly provided by Millipore.  

CaV3.2 - "High Throughput Pharmacology of CaV3.2 Channels on Nanion’s SyncroPatch 384PE"

icon sp96   SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) application note:   logo pdf   (0.6 MB)
Cells were kindly provided by Millipore.  

NaV1.5 - "High Throughput Pharmacology of NaV1.5 Channels on Nanion's SyncroPatch 384PE"

icon sp96   SyncroPatch 384i (a predecessor model of the SyncroPatch 384) application note   logo pdf   (1.5 MB)
Cells were kindly provided by Millipore.

TRPA1 - "High Throughput Activation and Block of hTRPA1 on Nanion’s SyncroPatch 384PE"

icon sp96   SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) application note   logo pdf   (0.6 MB)
Cells were kindly provided by Millipore.

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.

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.

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.

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

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

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.

hERG - "Characterization of hERG (CHO) on Nanion's Port-a-Patch"

icon pap   Port-a-Patch application note:   logo pdf   (0.6 MB)
Cells were kindly provided by Millipore.  

hERG - "Characterization of hERG (HEK293) on Nanion's Port-a-Patch"

icon pap   Port-a-Patch application note:   logo pdf   (0.7 MB)
Cells were kindly provided by Millipore.  

CaV3.2 - "Characterization of CaV3.2 on Nanion's Port-a-Patch"

icon pap   Port-a-Patch application note:   logo pdf   (0.7 MB)
Cells were kindly provided by Millipore.  

CaV2.2 - "Characterization of CaV2.2 on Nanion's Port-a-Patch"

icon pap   Port-a-Patch application note:   logo pdf   (0.6 MB)
Cells were kindly provided by Millipore.  

hERG - "Temperature controlled hERG recordings on the Port-a-Patch"

icon pap   Port-a-Patch application note:   logo pdf   (0.5 MB)
Cells were kindly provided by Millipore.  

hERG - "Pharmacology of hERG recorded on Nanion´s Port-a-Patch"

icon pap   Port-a-Patch application note:   logo pdf   (0.4 MB)
Cells were kindly provided by Millipore.  

NaV1.5 - Current Voltage Relationship

NaStrom IV

icon sp96   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.

 

 

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. 

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.

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.

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