NucGreen® Dead 488 ReadyProbes® Reagent Protocol for Fixed Cells
Nuclear stain for fixed cells
NucGreen® Dead 488 ReadyProbes® Reagent emits bright green fluorescence when bound to DNA and is suitable for staining nuclei in fixed-cell preparations and tissue sections. NucGreen® Dead 488 reagent may also be used to measure viability and to measure cytotoxicity in kinetic live-cell assays.
This protocol can be used for:
- Nucleic acid (nuclear) staining in fluorescence microscopy
This protocol should not be used for:
- Flow cytometry
You will need the following for this protocol:
- Cells growing in culture
- NucGreen® Dead 488 ReadyProbes® Reagent (Cat. No. R37109)
- Fluorescence microscope with FITC or GFP filters
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 NucGreen® Dead 488 Reagent per milliliter of medium.|
|3. Incubate for 5–15 minutes, protected from light.|
|4. Remove the stain solution.|
|5. Wash the cells 2–3 times in PBS.|
|6. Image the cells.|
|NucGreen® Dead 488|
|Standard filter set||FITC or GFP|
|EVOS® Light Cube||FITC|
|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).
HeLa cells stained with NucGreen® Dead 488 ReadyProbes® reagent, indicating loss of plasma membrane integrity.