Two-color nuclear staining assay for cell viability
This kit can be used to quickly and easily determine the viability of cells. NucBlue® Live reagent stains the nuclei of all cells, while propidium iodide stains only the nuclei of dead cells.
This protocol can be used for:
- Identifying live and 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
- ReadyProbes® Cell Viability Imaging Kit, Blue/Red (Cat. No. R37610)
- Fluorescence microscope with DAPI and RFP or TRITC filters
Protocol
 1. Culture cells in appropriate medium and vessel for microscopy |
 2. Add 2 drops each NucBlue® Live and propidium iodide per milliliter of medium to label cells |
 3. Incubate 5–30 minutes |
 4. Image cells |
Spectral information and storage
| NucBlue® Live | Propidium iodide | |
|---|---|---|
| Excitation/Emission | 360/460 nm | 535/617 nm |
| Standard filter set | DAPI | TRITC or RFP |
| EVOS® Light Cube | DAPI | TRITC or RFP |
| Storage conditions | Room temperature | 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 media with Live Cell Imaging Solution
Jurkat cells stained with NucBlue® Live reagent and propidium iodide.
For Research Use Only. Not for use in diagnostic procedures.
