NucGreen Dead 488 ReadyProbes Reagent | Thermo Fisher Scientific

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:

Cells growing in culture
NucGreen Dead 488 ReadyProbes Reagent (Cat. No. R37109)
Fluorescence microscope with FITC or GFP filters

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	504/523 nm
Standard filter set	FITC or GFP
EVOS Light Cube	FITC
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)

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