What microscope and assays do UK labs need to measure apoptosis and cell viability in living cells?
The question: “I need to see when my cells die — and what kind of death it is. What microscope and assay do I actually need?”
Apoptosis and viability imaging is one of the most common live-cell assays in drug discovery, toxicology and cancer biology. The right setup lets you distinguish early apoptosis from late apoptosis or necrosis, track the same cells over time, and quantify dose-response without fixing at every time point.
A Cell Death & Disease study (2016) showed that real-time high-content live-cell imaging gives robust kinetic apoptosis profiles, while the Assay Guidance Manual (NCBI Bookshelf) outlines standardised apoptosis marker assays for screening.
| Readout | What it reports | Typical probe / reporter | Imaging notes |
|---|---|---|---|
| Phosphatidylserine externalisation | Early apoptosis | Annexin V-AF488/AF647, pSIVA-IANBD | Needs Ca2+ in medium; avoid long light exposure |
| Membrane integrity loss | Late apoptosis / necrosis | PI, DRAQ7, SYTOX Green/Red, 7-AAD | Cell-impermeant dyes only enter compromised cells |
| Caspase activity | Execution-phase apoptosis | CellEvent Caspase-3/7 Green, NucView 488/530, FRET reporters | Genetic reporters best for repeated imaging; ratio imaging improves quantification |
| Mitochondrial membrane potential | Early apoptotic commitment | TMRM, JC-1, MitoTracker Red CMXRos | Sensitive to phototoxicity; use low LED power |
| Cell morphology / confluence | Viability and proliferation context | Transmitted light / phase / label-free segmentation | Complements fluorescence without extra dyes |
The EVOS M7000 inverted automated fluorescence microscope with OSI-2 Onstage Incubator is well suited to kinetic apoptosis assays in dishes or small multiwell formats. LED excitation is gentle enough for repeated time-lapse, and the environmental chamber keeps pH and temperature stable for hours to days.
For drug discovery and toxicology, the CellInsight CX5 and CX7 High Content Analysis platforms turn apoptosis assays into quantitative, plate-level readouts. With environmental control, they can also run live-cell kinetic screens.
| Use case | EVOS M7000 + OSI-2 | CellInsight CX5 / CX7 |
|---|---|---|
| Long kinetic films of the same cells | ✓ Ideal — low phototoxicity LED, stable environment | ✓ Possible with onstage incubator |
| Compound library screen | Manual or limited throughput | ✓ Purpose-built for multiwell HCA |
| Multi-parameter classification per cell | Open-source or Celleste analysis | ✓ HCS Studio with built-in apoptosis metrics |
| Cost and lab footprint | Benchtop, self-contained with OSI-2 compressor | HCS platform, larger footprint, higher throughput |
These references cover kinetic high-content apoptosis assays, membrane-based readouts, caspase imaging methods and standardised screening guidance.
Robust high-throughput kinetic analysis of apoptosis with real-time high-content live-cell imaging. A practical framework for plate-level time-course death assays.
View DOIBeyond annexin V: fluorescence response of cellular membranes to apoptosis. Reviews membrane-sensitive probes and their behaviour during programmed cell death.
View DOIHigh throughput ratio imaging to profile caspase activity, with potential application in multiparameter high-content apoptosis analysis and drug screening.
View DOIGuidelines for regulated cell death assays: a systematic summary and categorical comparison covering apoptosis, necroptosis and other death modalities.
View DOIImaging-based methods for assessing caspase activity in single cells, including fluorescent substrates and genetically encoded reporters.
View DOIAssay Guidance Manual chapter on apoptosis marker assays for high-throughput screening, summarising dye and reporter options, controls and data interpretation.
View NCBIPlatform pages for live-cell inverted imaging and high-content analysis, compatible with long-term apoptosis reporter time-lapse and multiwell screens.
EVOS M7000 Review CellInsight CX7 UKAn inverted fluorescence microscope with live-cell environmental control (37 °C, CO2, humidity) and multi-channel fluorescence is best. The EVOS M7000 with OSI-2 incubator works for long-term kinetic apoptosis imaging of dishes or small plates. For higher-throughput compound screens, the CellInsight CX5/CX7 High Content Analysis platform with environmental control is the practical choice.
Yes. Live-cell apoptosis assays use non-toxic membrane probes such as Annexin V-Alexa Fluor 488 or pSIVA, cell-impermeant dyes such as DRAQ7/SYTOX, and genetically encoded caspase reporters. These let you track when each cell enters early apoptosis, late apoptosis or necrosis over minutes to days.
Early apoptosis is usually detected by phosphatidylserine externalisation (Annexin V binding) and caspase activation. Late apoptosis / secondary necrosis is marked by loss of membrane integrity, allowing cell-impermeant nuclear dyes such as propidium iodide or DRAQ7 to enter. Combining both readouts helps distinguish apoptosis from necrosis.
They report different things. Annexin V detects phosphatidylserine flip, an early membrane change. Caspase reporters report protease activity inside the apoptotic execution pathway. Using both gives a more complete kinetic picture of cell death.
Maintain temperature, CO2 and humidity with an onstage incubator. Use low phototoxicity LED excitation, minimal laser power if confocal, and short exposure times. For multi-day studies, refresh medium or use a perfusion chamber if the assay requires it.
Yes. CellInsight CX5/CX7 can image 96- or 384-well plates, segment individual cells, quantify Annexin V or caspase reporter intensity, count viable/dead cells, and export dose-response data for drug candidates.