Nuclear stain for fixed cells
NucBlue Fixed Cell ReadyProbes Reagent is a bright blue cell-impermeant nuclear stain for fixed cells. It is useful to distinguish the condensed nuclei of apoptotic cells. It is also suitable for staining nuclei in fixed-cell preparations and tissue sections.
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 Fixed Cell ReadyProbes Reagent (Cat No R37606)
- Fluorescence microscope with UV light source and a blue/cyan emission filter
Protocol
Labeling fixed cells
First, fix and permeabilize cultured cells with a protocol appropriate for your sample.
 1. Wash the cells 1–3 times in PBS as needed. |
 2. Add two drops of NucBlue Fixed Cell ReadyProbes Reagent per milliliter of medium. |
 3. Incubate for 5 minutes at 20–25°C. |
 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).
Fixed U2OS cell staining. U2OS cells were fixed and permeabilized using the Image-iT Fixation/Permeabilization Kit. Cells were then with NucBlue Fixed Cell ReadyProbes Reagent for 15 minutes and imaged.