Membrane integrity-based viability assay
NucGreen Dead 488 ReadyProbes Reagent is a cell-impermeant stain that emits bright green fluorescence when bound to DNA. Since this reagent stains cells that lack plasma membrane integrity, it is extremely useful to measure viability and to measure cytotoxicity in kinetic live-cell assays. NucGreen Dead 488 reagent is also suitable for staining nuclei in fixed-cell preparations and tissue sections.
This protocol can be used for:
- Identifying dead cells using a fluorescence microscope
This protocol should not be used for:
- Flow cytometry
You will need the following for this protocol:
- Cells growing in culture
- NucGreen Dead 488 ReadyProbes Reagent (Cat. No. R37109)
- Fluorescence microscope with FITC or GFP filters
Protocol
 1. Culture cells in an appropriate medium and vessel for fluorescence microscopy |
 2. Add 2 drops NucGreen Dead 488 reagent per milliliter of medium |
 3. Incubate for 5 minutes at 20–25°C |
 4. Image the cells |
Spectral information and storage
| NucGreen Dead 488 | |
|---|---|
| Excitation/Emission | 504/523 nm |
| Standard filter set | FITC or GFP |
| EVOS Light Cube | FITC |
| Storage conditions | Room temperature |
Protocol tips
- In some cases, more or fewer drops may be needed to achieve optimal staining intensity
- Image quality may be improved by replacing the culture medium with Live Cell Imaging Solution (Cat. No. A14291DJ)
HeLa cells stained with NucGreen Dead 488 ReadyProbes reagent, indicating loss of plasma membrane integrity.