NucBlue® Fixed Cell ReadyProbes® Reagent Protocol
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
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.
|Standard filter set||DAPI|
|EVOS® Light Cube||DAPI|
|Storage conditions||Room temperature|
- 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).