CardioExcyte 96 – Combined contractility, electrophysiology and cell viability

The CardioExcyte 96 is a hybrid system recording both contractility and electrophysiology of intact cardiomyocyte networks. In addition, the base impedance is continuously and automatically monitored as a measure of acute and chronic cell activity. No subtleties of cytotoxic responses are missed, this includes non-contractile cell types such as hepatocyte-like cells or cancer cells as well as contractile cardiac cells. Impedance and extracellular field potential measurements are performed at high resolution, are non-invasive and label-free. The CardioExcyte 96 is a fully automated device, recording from 96 wells at a time. The benefits the CardioExcyte 96 offers include:

Impedance & extracellular field potentials from the same well
96 recording wells in parallel
Non-invasive & label-free measurements
High resolution
Electrical and optical pacing options
Acute and chronic assessment of cell activity and toxicity
Part of the CiPA validation study

With embedded electronics and a sophisticated sensor technology integrated in the well-plate based consumables, the CardioExcyte 96 is a turn-key system for efficient impedance and extracellular field potential measurements. The CardioExcyte 96 package includes an Incubation System which makes the system independent of an incubator. The system is placed on the bench top with maintained control over temperature, gas mix and humidity.

Efficiency and ease-of-use are facilitated by an outstanding software package for data handling and export - an essential part of the CardioExcyte 96 system. The system has been validated with stem cell-derived cardiomyocytes from all major stem cell providers, as monolayers or beating 3D-clusters, as well as with cell lines (hepatocyte-like cells, cancer cells etc.) for cytotoxicity studies.

For detailed information:

Add-Ons and Features

CardioExcyte 96 Platform

CardioExcyte Integra 1

The CardioExcyte 96 package contains an Integra Viaflo Assist that provides automated multichannel pipetting for reproducible 96 well plate handling of cells and compounds.

CardioExcyte 96 Incubation System

CardioExcyte Incubation System 1

The CardioExcyte 96 package includes an Incubation System. This means that the measurements are performed outside the cell culture incubator and the complete setup fits on a lab benchtop. The CardioExcyte 96 Incubation System controls temperature, gas mix and humidity.

CardioExcyte 96 SOL (Add-On)

Optical stimulation and investigation of impedance and extracellular field potentials in parallel:

CardioExcyte Sol 1

The stimulating optical lid, CardioExcyte 96 SOL, uses LEDs for spatially uniform stimulation of cells transfected with light-gated ion channels such as Channelrhodopsin2 (ChR2). After transfection of iPS cardiomyocytes with ChR2 the cells can be stimulated by blue light. Each light impulse induces a depolarization of the cells and thus an action potential is triggered. The advantages of optical stimulation over electrical stimulation include the highly precise timing, all cells of the beating network are stimulated exactly at the time of the light stimulus. In contrast, electrical stimulation propagates from the electrode across the well and thus cells are stimulated progressively. A mean beat calculation of precisely timed beats enables in-depth compound analysis and concentration response dependencies to be obtained. This is one of the key software capabilities of the CardioExcyte Control/ DataControl software package that is provided with each CardioExcyte 96 system.

Software

CardioExcyte 96 Software package

CardioExcyte Analysis Software

The CardioExcyte Control software supports online analysis of beating parameters. The unique Mean Beat Function automatically visualizes the average of beat traces from one well, enveloped by the standard deviation, indicating the consistency of the beat pattern within the recording well, i.e. if the cellular network is synchronously beating or not. The control software is intuitive and easy-to-use, at the same time very powerful in terms of experimental setup and online visualization of the recorded data.

The DataControl 96 software package is an independent software package to load and analyze data recorded on the CardioExcyte 96.The analysis template can be saved and reloaded. Customized export formats of analysis results are integrated.

The Online Analysis Post Processor (OAPP) is a software tool enabling quick visualization of well-to-well statistics and concentration dependent effects. Parameters and traces can be normalized to control values or time points. Export of graphics is straightforward making OAPP a highly versatile tool for the analysis routine.

Statement about the DataControl 96 Package

Hans-Peter Scholz - Statement about the Analysis Software of the CardioExcyte 96 "DataControl 96"

Icon CE “DataControl 96 allows us to analyze large amounts of data and represents a huge step towards standardized experiment analysis. All the well's individual raw data, analysis results, concentration response curves and also the fits averaged per compound are displayed on a single screen, which means that all relevant information is right at the place where we want to see it. Analysis templates can be used to re-do the same analysis procedures on a different compound, but same target, thus reducing the effort for data handling to just a couple of mouse clicks. Powerful averaging with standard deviation display and referencing methods allow for great quality control. Furthermore, multiple export options are available to allow a seamless integration in databases.”

Hans-Peter Scholz, Technician,
Biopharma, Global Early Non Clinical Safety, Merck KGaA, Darmstadt, Germany

Consumables

NSP-96

CE Plate

The NSP-96 chip is a proprietary innovative product by Nanion Technologies, developed for the CardioExcyte 96. It is produced and quality-assured partly by an external partner and partly in-house at Nanion headquarters and shipped from Munich to our international customers. Different types of NSP-96 chips are available for different experiments.

Description and Material

Standard 96-well plate format
Working volume: 25-340 µl (surface 34mm²)
Planar, bipolar circular gold electrodes
Supports recordings from 2D and 3D structures
Hybrid sensor design for both EFP and impedance recordings

Available chip types

"NSP-96, CardioExcyte 96 Sensor Plates - Stim": Type standard with extra stimulation electrode (Order # 201003)
"NSP-96 CardioExcyte96 Sensor Plates 0.6 mm": Type standard, 0.6 mm recording electrode (Order # 201002)
"NSP-96 CardioExcyte96 Sensor Plates 2.0 mm": Type standard, 2.0 mm recording electrode (Order # 201001)

Other types are available upon request!

Testimonials & Case Studies

Prof. Bjorn C. Knollmann - Statement about the CardioExcyte 96

Icon CE “We are happy to have acquired the CardioExcyte 96 from Nanion and look forward to publishing the first of many papers. We looked at several different devices on the market but were impressed by the resolution and ease-of-use of the CardioExcyte 96 and Nanion’s enthusiasm and friendly customer support. I was particularly intrigued by the pacing feature, the single electrode design giving better signal to noise ratio, the dual EFP + impedance capability, and the environmental chamber so that you can run everything on the benchtop. The new optical pacing feature added in 2017 has further expanded the utility of the instrument for drug testing and disease modeling: For the first time, rate-dependent changes of cardiac electrophysiology and contractility can be easily quantified.”

Prof. Bjorn C. Knollmann, Director of the Center for Arrhythmia Research and Therapeutics of Vanderbilt University School of Medicine:

Marc Rogers - Statement about the Patchliner and CardioExcyte 96

"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

Hans-Peter Scholz - Statement about the Analysis Software of the CardioExcyte 96 "DataControl 96"

Hans-Peter Scholz, Technician,
Biopharma, Global Early Non Clinical Safety, Merck KGaA, Darmstadt, Germany

Dr. Ralf Kettenhofen - Statement about the CardioExcyte 96

Icon CE "CardioExcyte 96 is an easy-to-use system, providing impedance-based and MEA-like cardiac safety data from a diversity of stem cell-derived cardiomyocytes, and constitutes an excellent complement to automated patch clamp-based safety screening. Concentration- and time-dependence of a compound’s potential cardiotoxicity can efficiently be obtained where the alteration of beating patterns can give a hint as to which cardiac ion channel(s) is affected, after which detailed electrophysiology investigations can be undertaken. Cardiac network responses offer a comprehensive view of a compound’s safety profile, without having to use in-vivo methods, which saves time, costs and suffering. Further on, the powerful software, used for recordings and analysis, employs comprehensive beat investigation algorithms, displaying detailed beating kinetics in real-time. Data handling and export is straightforward, easy to grasp and yet very, very powerful."

Dr. Ralf Kettenhofen, Head of Laboratory
Ncardia, Cologne, Germany.

Dr. Daniel Konrad - Statement about the CardioExcyte 96

Icon CE “Impedance and field potential data of beating cardiomyocytes is getting increasingly important to obtain a comprehensive view of the cardiac liability of test compounds. To answer these questions for our clients, B’SYS successfully implemented the CardioExcyte 96 system and provides related validated services to its customers, mostly using ipS derived cardiomyocytes. B’SYS’ scientists are pleased with the CardioExcyte’s ease of operation and data acquisition. The powerful software allows for applying meaningful data analysis and time saving export of data as needed for our daily customer reports.“

Dr. Daniel Konrad
CEO B'SYS

Anika Duenbostell-Schmidt - Statement about the CardioExcyte 96

Icon CE "I use the CardioExcyte 96 weekly to test substances on our human iPSC-derived cardiomyocytes. It allows me to map, compare and evaluate a wide variety of pharmacological responses with a high degree of reliability and uniformity, and evaluate them through the rapid analysis function."

Anika Duenbostell-Schmidt, Head QC at Ncardia
Ncardia, Cologne, Germany.

Data and Applications

NCX1 - Effect of block of cardiac NCX on beat rate

Icon CE CardioExcyte data and applications:
Cells were kindly provided by Cellular Dynamics.

Block of NCX1 expressed in iCell^® Cardiomyocytes² by SEA 0400 caused a dose dependent increase in beat rate.

Cardiac Ion Channels - Pharmacology of Vandetanib

Icon CE CardioExcyte 96 and 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, Na_V1.5, Ca_V1.2 and K_V4.3. 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.

Cardiac Ion Channels - Pharmacology of Sotalol

Icon CE CardioExcyte 96 and SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) 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.

Cardiomyocytes - Data from different cell providers

Icon CE CardioExcyte data and applications:
Cells were kindly provided by Axiogenesis, Cellular Dynamics, GE Healthcare, Pluriomics, ReproCell, Takara Bio Cellartis Clonetech.

The CardioExcyte 96 allows for
• Non-invasive, label-free measurements of beating cardiomyocyte networks
• 96 recording wells in parallel with 1 ms time resolution
• Quick experiments or long-term compound effects on cardiotoxicity
• Real-time access to beating parameters
• Outstanding software for data analysis and export
• Cost effcient consumables - 96-well format

Cardiomyocytes - Time-dependent effect of Pentamidine on Cor.4U cardiomyocytes

Icon CE CardioExcyte data and applications:
Cells were kindly provided by Axiogenesis.

Time-dependent effect of Pentamidine on Cor.4U cardiomyocytes. Pentamidine clinically causes acquired long QT syndrome, which is associated with prolonged QT intervals, tachycardias, and sudden cardiac arrest. Pentamidine delays terminal repolarization in human heart by acutely blocking cardiac inward rectifier currents. At the same time, it reduces surface expression of the cardiac potassium channel IKr/human ether à-go-go-related gene (hERG).
Insets: EFP raw traces and online analysis value FPDc in the presence of Pentamidine (10 µM) at different timepoints.

Cardiomyocytes - Tetracaine dose response curves as recorded with Cor.4U cells

Icon CE CardioExcyte data and applications:
Cells were kindly provided by Axiogenesis.

Impedance amplitude is not changed by addition of increasing concentrations of Tetracaine (left panel), while beat rate of Cor.4U^® cells is decreasing. For example, 29.6µM of Tetracaine decreased the beat rate by ~60% when compared to pre-addition values. Cumulative dose-response relationships indicate Tetracaine potency for same-well additions. Representative raw traces for impedance signals (middle panel) clearly indicate a decrease in cell monolayer beat rate with increasing concentrations of Tetracaine.
Extracellular Field Potential (EFP) spike amplitude is decreased by cumulative Tetracaine dose applications to the same monolayer of Cor.4U^® cardiomyocytes (top right), in agreement with compound mechanism of action. Representative raw traces for EFP signals (bottom graph) clearly indicate a decrease in spike amplitude.

Cardiomyocytes - Nifedipine and its concentration dependent effect on Cor.4U cells

Icon CE CardioExcyte data and applications:
Cells were kindly provided by Axiogenesis.

Nifedpinie is a dihydropyridine calcium channel blocker that primarily blocks L-type calcium channels. Impedance and EFP recordings on Cor.4U cells reveal a concentration dependent effect on impedance amplitude, beat rate and also a shortening of the FPD as expected.

Cardiomyocytes - Effect of E4031 on the impedance and EFP signals on iCell cardiomyocytes

Icon CE CardioExcyte data and applications:
Cells were kindly provided by Cellular Dynamics.

Effect of the specific hERG blocker, E4031, on the impedance and EFP signals. A Impedance signal of 12 wells in control conditions (left) and the same 12 wells after 13 mins incubation in E4031 at the concentrations indicated (right). E4031 (300 nM, 3 mins incubation) induces EAD, shown in the inset, which can lead to potentially fatal ventricular arrhythmias. B EFP signal of 12 wells in control conditions (left) and the same 12 wells after 13 mins incubation in E4031 at the concentrations indicated (right). E4031(300 nM, 30 mins incubation) also causes arrhythmic effects in the EFP mode shown in the inset.

Cardiomyocytes - Effect of the ß-adrenergic receptor agonist, Isoproterenol, on the impedance and EFP signals on iCells

Icon CE CardioExcyte data and applications:
Cells were kindly provided by Cellular Dynamics.

A Impedance signal of 12 wells in control conditions (left) and the same 12 wells after 13 mins incubation in isoproterenol at the concentrations indicated. Isoproterenol acts as a heart stimulant, it increases heart rate, an effect which is highlighted in the inset where the trace in the presence of isoproterenol (300 nM; black) and the control trace (red) are shown overlaid. In the presence of even low concentrations of isoproterenol (1 nM), an increase in the beat rate is observed. B Bar graph showing beat rate before (dark blue) and after (light blue) incubation in isoproterenol at the concentrations indicated. At all concentrations, a clear increase in beat rate is observed. C EFP signal of 12 wells in control conditions (left) and the same 12 wells after 30 mins incubation in isoproterenol at the concentrations indicated (right). Isoproterenol shortens the FPD, an effect which is highlighted in the inset where the trace in the presence of isoproterenol (300 nM; black) and the control trace (red) are shown overlaid.

Cardiomyocytes - Effects of verapamil on impedance (left) and EFP (right) signals of Cellartis Cardiomyocytes

Icon CE CardioExcyte data and applications:
Cells were kindly provided by Takara Bio Cellartis Clontech.

(A) Impedance (left) and EFP signals (right) in control conditions
(B) Upon application of 10 nM verapamil
(C) Upon application of 100 nM verapamil.
(D) The mean beats for impedance (left) and EFP (right) are shown in D

Cardiomyocytes - Effects of sotalol on impedance (left) and EFP (right) signals of Cellartis Cardiomyocytes

Icon CE CardioExcyte data and applications:
Cells were kindly provided by Takara Bio Cellartis Clontech.

Impedance (left) and EFP signals (right) in control conditions (A) and 1 μM sotalol (B) are shown.
Irregular beating can be observed in the presence of sotalol.

Murine Mammary Carcinoma Cells - Effect of CAF (cyclophosphamide, adriamycin (doxorubicin) and 5-fluouracil)

Icon CE CardioExcyte data and applications:
Data kindly provided by Oliver Reinhardt, MPI Experimentelle Medizin, Göttingen

Murine mammary carcinoma cells (H8N8 and H8N8 T3.2) were monitored over a time period of 500 h. Changes in impedance, and therefore confluency, were used as a measure of toxicity.
H8N8 cells have been developed as a tool to investigate tumor cell recurrence, chemoresistance and epithelial-to-mesenchymal transition (EMT). Cells were treated with increasing concentrations of CAF (cyclophosphamide, adriamycin (doxorubicin) and 5-fluouracil) on the CardioExcyte 96 during impedance measurements. Therefore, the CardioExcyte 96 in combination with murine mammary carcinoma cells provides a novel tool for investigating therapy resistance of cancer cells in vitro.
Data kindly provided by Oliver Reinhardt, MPI Experimentelle Medizin, Göttingen

Cardiomyocytes - Optogenetics meets cardiac safety

Icon CE CardioExcyte data and applications:
Cells were kindly provided by Axiogenesis.

The stimulating optical lid, CardioExcyte 96 SOL, uses LEDs for spatially uniform stimulation of cells transfected with light-gated ion channels such as Channelrhodopsin2 (ChR2).

Right graph: LPM – Light pulse per minute plotted against the recorded beat rate (average of 96 wells). ChR2 transfected Cor.4U cells are following the optical pace rate.

Cardiomyocytes - Channelrhodopsin 2 (ChR2) transfected Cor.4U cells and optical pacing

Icon CE CardioExcyte data and applications:
Cells were kindly provided by Axiogenesis.

ChR2 transfected Cor.4U cells are following the optical pace rate.

Raw data traces upon a 1 Hz, 1.5 Hz, 2 Hz., 2.5 Hz and 3 Hz stimulation rate, extracellular field potentials (top) and impedance (bottom).

Cardiomyocytes - Myocyte phase II study: CiPA conform analysis and arrhythmia detection

Icon CE CardioExcyte data and applications:
Cells were kindly provided by Axiogenesis.

Nanion developed a CiPA conform analysis for the Myocyte phase II study. The feature comes along is included in our CiPA analysis routine. Automated arrhythmia detection is just one highlight out of many when it comes to the CardioExcyte 96 software.

Hepatocytes - Paracetamol toxicity on hepatocyte-like MetaHeps cells

Icon CE CardioExcyte data and applications:
Cells were kindly provided by MetaHeps.

The impedance signal recorded on the CardioExcyte 96 changes as a result of alterations in confluency, cell contact (morphological shape) and conductivity of adherent cells and thereby provides a measure of toxicity. MH cells from MetaHeps^® were grown on NSP-96 plates and base impedance was monitored over time. Increasing concentrations of paracetamol induce a decrease in base impedance of MH cells which can be monitored continuously. Tween (2%) induced 100% cell death and was used as a positive control.

Webinars and Movies

Webinars

12.09.2018 | Webinar: CiPA study: Bridging ion channel and myocyte data

180912 event image CiPAII Webinar

Icon CE CardioExcyte 96, Patchliner and SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) Webinar

Date: September 12, 4:00 PM CEST (10:00 AM EDT)

Get up-to-date with the CiPA progress of the Myocyte and Ion Channel Work Goups:

CiPA myocyte phase II validation study results: cross-site comparison using the CardioExcyte 96
HTS Phase I study: an update on progress of the CiPA Ion Channel Work Stream using the SyncroPatch 384PE and Patchliner

28.04.2015 | Webinar: Excited About Contraction – Combining Contractility and Excitability Measurements in Cardiotoxicity Screening

Icon CE CardioExcyte 96

This Webinar covers the introduction of the CardioExcyte 96, a hybrid device for impedance- and MEA-type recordings from intact, beating networks of stem cell-derived cardiomyocytes.

Movies: Tutorials and Oral Presentations

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 Icon CE 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 - CiPA myocyte phase II validation study results: cross-site comparison using the CardioExcyte 96

Icon CE CardioExcyte 96 Oral Presentation

Presenter:
Dr. Sonja Stölzle-Feix, Nanion Technologies
Source:
Webinar: "CiPA study: Bridging ion channel and myocyte data", September 12, 2018

2017 - Impedance-Based Monitoring of Cell-Based Assays: The Power of MultiFrequency Recordings

Icon CE CardioExcyte 96 Oral Presentation

Presenter:
Prof. Dr. Joachim Wegener, University of Regensburg, Germany

2017 - CardioExcyte 96 Tutorial Video

Icon CE CardioExcyte 96

Short introduction on the CardioExcyte 96 (tutorial video, 3 minutes)

Downloads:

Application Notes

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

SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) Patchliner Icon CE CardioExcyte 96 application note (2.3 MB)

CardioExcyte 96

Combined impedance and MEA-like recordings

CardioExcyte 96

For cardiac safety screening

CardioExcyte 96

Next generation label-free cell analysis

CardioExcyte 96

Intuitive data analysis & arrhythmia detection

CardioExcyte 96

Transparent plates available for imaging

CardioExcyte 96 – Combined contractility, electrophysiology and cell viability

For detailed information:

Add-Ons and Features

CardioExcyte 96 Platform

CardioExcyte 96 Incubation System

CardioExcyte 96 SOL (Add-On)

Software

CardioExcyte 96 Software package

Statement about the DataControl 96 Package

Consumables

NSP-96

Description and Material

Available chip types

Testimonials & Case Studies

Data and Applications

Webinars and Movies

Webinars

Movies: Tutorials and Oral Presentations

Downloads:

Application Notes

Product Sheets

Publications

Posters

Presentations