Membrane integrity–based viability assay

NucGreen® Dead 488 ReadyProbes® Reagent is a cell-impermeant stain that emits bright green fluorescence when bound to DNA. Since this reagent stains cells that lack plasma membrane integrity, it is extremely useful to measure viability and to measure cytotoxicity in kinetic live-cell assays. NucGreen® Dead 488 reagent is also suitable for staining nuclei in fixed-cell preparations and tissue sections.

This protocol can be used for: 

  • Identifying dead cells using a fluorescence microscope

This protocol should not be used for: 

  • Flow cytometry

You will need the following for this protocol:

1. Culture cells in an appropriate medium and vessel for fluorescence microscopy. 
2. Add 2 drops NucGreen® Dead 488 reagent per milliliter of medium.
3. Incubate for 5 minutes at 20–25°C.
4. Image the cells.

Spectral information and storage

  NucGreen® Dead 488
Excitation/Emission (nm) 504/523  
Standard filter set FITC or GFP
EVOS® Light Cube FITC
Storage conditions Room temperature


Protocol tips

HeLa cells stained with NucGreen® Dead 488 ReadyProbes®
HeLa cells stained with NucGreen® Dead 488 ReadyProbes® reagent, indicating loss of plasma membrane integrity.