This article and associated images are based on a poster originally authored by Denise Sullivan, Zhong Yu, Benjamin Streeter, Stacie Chvatal and Daniel Millard and presented at ELRIG Drug Discovery 2024 in affiliation with Axion BioSystems.
This poster is being hosted on this website in its raw form, without modifications. It has not undergone peer review but has been reviewed to meet AZoNetwork's editorial quality standards. The information contained is for informational purposes only and should not be considered validated by independent peer assessment.
Maestro Z: Dynamic cell tracking
Impedance technology
Cell-based in vitro cytotoxicity assays are valuable tools for screening compounds for toxicity evaluation. Many in vitro cytotoxicity assays rely on dyes or labels to measure cell death at a single time point after a predetermined exposure time.
Axion BioSystems’ Maestro Z platform offers impedance-based cell analysis for real-time, label-free monitoring of cell viability, morphology, cytolysis, and signaling. We used the Maestro Z to characterize a cytotoxicity assay for high-throughput screening and dose-response analysis.
Image Credit: Image courtesy of Denise Sullivan et al., in partnership with ELRIG (UK) Ltd.
The impedance is measured from electrodes embedded in the bottom of each well. As cells cover more of the electrode, impedance increases proportionately to the number of viable cells. If a perturbation kills the attached cells, impedance decreases as the cells lyse.
The Maestro Z product family
Image Credit: Image courtesy of Denise Sullivan et al., in partnership with ELRIG (UK) Ltd.
- Label-free, non-invasive tracking of cultured cells or spheroids/organoids
- Integrated environmental control provides a stable benchtop environment for short- and long-term toxicity studies
- Automatic and continuous cell monitoring from 96 or 384 wells simultaneously
- “One button setup” automatically docks the plate and adjusts temperature and CO2 levels
- Powerful data analysis to focus on the science, while AxIS Z handles the details with simple setup and automatic experiment tracking
- See your cells with the viewing window included in each well of the CytoView-Z 96-well plate.
- State-of-the-art electrode processing chip (BioCore v4) offers stronger signals, ultra-low frequency content, and enhanced flexibility
Image Credit: Image courtesy of Denise Sullivan et al., in partnership with ELRIG (UK) Ltd.
Real-time, label-free cytotoxicity assay
Impedance assay measures diverse cell properties
The Maestro Z records impedance at multiple frequencies simultaneously, enabling a thorough characterization of cell behavior, including:
- Coverage/Density—The change in impedance is directly related to the quantity of cells in a 2D and 3D culture covering the electrodes.
- Cytotoxicity – Dynamic monitoring of cell viability measures the degree and speed of cell death.
- Signaling – Small changes in cell shape or cytoskeleton organization are detected in response to intracellular signaling events.
Image Credit: Image courtesy of Denise Sullivan et al., in partnership with ELRIG (UK) Ltd.
Dose-response analysis for Maestro Z cytotoxicity assay
Dose response analysis quantifies the relationship between the dose delivered and a specific biological effect. Dose response analysis classically utilizes the Hill equation model which can incorporate the physical constraints of the system (e.g. Cytolysis must be between 0% and 100%) and provides an estimate of the dose producing the half-maximal response (IC50 or EC50).
Image Credit: Image courtesy of Denise Sullivan et al., in partnership with ELRIG (UK) Ltd.
Image Credit: Image courtesy of Denise Sullivan et al., in partnership with ELRIG (UK) Ltd.
- SKOV3 cells were treated with doxorubicin in a 9-point concentration sweep. Resistance (above, left) and cytolysis (above, middle) were plotted over time.
- Dose-response analysis (above, right) measures the EC50 of doxorubicin at 0.4 μM using a Hill equation fit.
- Dose-response analysis was performed for five different drugs in a separate assay using a CytoView-Z 384-well plate and A549 target cells. The EC50 is displayed for each of the five drugs.
Multi-plate dose-response analysis
Multi-plate CAR T cell potency dose response with the TrayZ
The TrayZ platform can simultaneously perform dose-response analysis from four CytoView-Z 96-well plates with label-free impedance measurements inside a standard incubator. Here, we show the EC50 for Paclitaxel and Doxorubicin, dosed onto SKOV3 cells, across four plates measured simultaneously. We observed strong agreement in the dose-response analysis across plates for each compound.
Image Credit: Image courtesy of Denise Sullivan et al., in partnership with ELRIG (UK) Ltd.
Image Credit: Image courtesy of Denise Sullivan et al., in partnership with ELRIG (UK) Ltd.
- KOV3 cells were treated with paclitaxel and doxorubicin (left) in a 6-point concentration sweep.
- Dose-response analysis (above, right) measures the EC50 consistently across plates.
Scalable dose-response analysis with TrayZ cytotoxicity assay
Not only can the TrayZ platform be used to study the dose-response of anti-tumorigenic compounds, but it also can be used to study the cytotoxic potential of chimeric antigen receptor (CAR) T cells delivered at multiple E:T ratios. Here, we cocultured HER2-positive SKOV3 target cells treated with CAR T cells targeted toward the HER2 antigen.
We measured SKOV3 cytolysis caused by the CAR T cells at various E:T ratios (1:5, 1:2, and 2:1). We also analyzed the Kill Time 50 (KT50) values (i.e., the time it took each CAR T cell dose to induce 50% cytolysis) and found consistent results across all four plates measured on the TrayZ.
Image Credit: Image courtesy of Denise Sullivan et al., in partnership with ELRIG (UK) Ltd.
Image Credit: Image courtesy of Denise Sullivan et al., in partnership with ELRIG (UK) Ltd.
- SKOV3 cells were dosed with HER2-targeted CAR T cells at three different E:T ratios, and cytolysis was measured in real-time over multiple days.
- Dose-response analysis (above, right) measured the KT50 values consistently across all four plates on the TrayZ system.
About Axion BioSystem
Axion BioSystems’ suite of live-cell analysis tools offers next-generation solutions for continuous, noninvasive monitoring of in vitro cell models on your benchtop or inside your incubator. From investigating disease mechanisms in neurology and cardiology to developing lifesaving immunotherapies and antivirals, Axion’s award-winning products are designed to accelerate scientific breakthroughs and transform therapeutic development from discovery to manufacturing.
About ELRIG (UK) Ltd.
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