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NucBlue Live ReadyProbes Reagent Protocol
Nuclear stain for live cells
NucBlue Live reagent (Hoechst 33342 dye) stains the nuclei of all cells. NucBlue Live reagent is frequently used to distinguish condensed nuclei in apoptotic cells.
This protocol can be used for:
- Nuclear staining to identify nuclear condensation in apoptotic cells using fluorescence microscopy
This protocol should not be used for:
- Flow cytometry
You will need the following for this protocol:
- Cells growing in culture
- NucBlue Live ReadyProbes Reagent (Cat. No. R37605)
- Fluorescence microscope equipped with a DAPI filter
Protocol
 1. Culture cells in an appropriate medium and vessel for fluorescence microscopy. |
 2. Add two drops of NucBlue Live ReadyProbes Reagent per milliliter of medium. |
 3. Incubate for 20 minutes, protected from light. |
 4. Image the cells. |
Spectral information and storage
| NucBlue | |
|---|---|
| Excitation/Emission (nm) | 360/460 |
| Standard filter set | DAPI |
| EVOS Light Cube | DAPI |
| 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).
U2OS live-cell staining. Live U2OS cells were imaged 15 minutes following staining with NucBlue Live ReadyProbes Reagent.