Membrane integrity—based viability assay
This kit includes Image-iT® DEAD Green™ viability stain for identification of dead cells. After staining your cells, the signal from the stain will be retained if the sample is fixed and permeabilized.
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
- Image-iT® DEAD Green™ Kit (Cat. No. I10291)
- Phosphate-buffered saline (Cat. No. 14190-144)
- Anhydrous DMSO (Cat. No. D12345)
- Fluorescence microscope with FITC filters
Protocol
 1. Culture cells in appropriate medium and vessel for microscopy |
 2. Dilute Image-iT® DEAD Green™ stain by adding 225 µL DMSO to vial to make a 1,000X stock solution |
 3. Add 1 µL stock solution per milliliter of medium to label cells |
 4. Incubate 30 minutes at 37°C |
 5. Image cells |
Spectral information and storage
| Image-iT® DEAD Green™ | |
|---|---|
| Excitation/Emission | 488/515 nm |
| Standard filter set | FITC or GFP |
| EVOS® Light Cube | GFP |
| Storage conditions | Room temperature |
Protocol tips
- This kit is compatible with antibody labeling protocols
- Dye stock solutions should be stored frozen
Valinomycin-treated HeLa cells labeled with the Image-iT® DEAD Green™ Kit.
