ReadyProbes® Cell Viability Imaging Kit (Blue/Red) Protocol

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.
| Resources | ||
For Research Use Only. Not for use in diagnostic procedures.
 
                     
                     
                  
                 
        
         
    








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