Real-Time Apoptosis Assays
Apoptosis is a programmed and regulated process of cell death where cells activate specific intrinsic pathways that cause them to shrink, condense, and eventually be cleared by phagocytosis. Relative to necrotic cell death where cell membrane integrity is compromised, leading to cell injury and inflammation, apoptotic cell death is less destructive and limits disruption of nearby cells and tissue. Typical biomarker-based assays require multiple handling steps and only yield endpoint measurements.
Explore Agilent xCELLigence real-time cell analysis technologies to learn about a more comprehensive apoptosis assay that provides:
Direct measurement of cell death and behavior
Quantitative and reproducible apoptotic data
A simple and high-throughput method
Detailed insight into cellular mechanism of action
Multiple Perspectives into Apoptosis to Increase Confidence
From the exact same population of cells, gain deeper insights into drug-mediated apoptosis with xCELLigence RTCA eSight. Automatically quantify and visualize apoptosis over the course of multiple days, without additional sample processing steps. Capture early and late apoptotic events such as modulation of cell adhesion, cell membrane integrity, and cell detachment. Simultaneous real-time cellular impedance and live cell imaging provide sensitive information about cell viability, growth, and morphology.
Watch Video
Multiplex Impedance-Based Data with Live Cell Imaging
While the standard xCELLigence RTCA instruments (SP, MP, HT models) provide comprehensive, noninvasive and label-free analysis in 96- or 384-well formats, the xCELLigence RTCA eSight adds live cell imaging for increased confidence in your cell analysis and conclusions. In one instrument, capture five different perspectives (impedance, brightfield, red, green, and blue fluorescence imaging) on the exact same population of cells to reveal greater insight into cell health.
Visualize and Quantify Apoptosis with Agilent eSight Reagents
With three distinct fluorescence channels and brightfield imaging, xCELLigence RTCA eSight provides provides both qualitative and quantitative assessment of cell morphology, size, and confluence under various assay conditions.Combine fluorescent live-imaging reagents and dyes, such as Agilent eAnnexin V, eCaspase-3, and eLenti, for multiplexed measurements of cell viability, apoptosis, and cytotoxicity in a single well. Additionally, genetically encoded, pathway-specific fluorescent reporters are also compatible.
Determine Effective Drug Concentrations from Multiple Readouts to Confirm Accuracy
Generate dose-response curves from impedance and image-based readouts to verify the concentration of a drug that gives half-maximal response (EC50). In this example, the area under the curve, spanning from the time of drug addition to 60 hours after drug addition, can be plotted as a function of drug concentration to yield the dose-response curves. The four different readouts resulted in R2 values ranging from 0.96 to 0.98 and EC50 values from 0.86 to 3.0 µm.