Two-color nuclear staining assay for cell viability

This kit can be used to quickly and easily determine the viability of cells. NucBlue® Live reagent stains the nuclei of all cells, while propidium iodide stains only the nuclei of dead cells.

This protocol can be used for:

  • Identifying live and dead cells using a fluorescence microscope

This protocol should not be used for:

  • Flow cytometry

You will need the following for this protocol:

Protocol

1. Culture cells in appropriate medium and vessel for microscopy
2. Add 2 drops each NucBlue® Live and propidium iodide per milliliter of medium to label cells
3. Incubate 5–30 minutes
4. Image cells

Spectral information and storage

  NucBlue® Live Propidium iodide
Excitation/Emission 360/460 nm 535/617 nm
Standard filter set DAPI TRITC or RFP
EVOS® Light Cube DAPI TRITC or RFP
Storage conditions Room temperature 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 media with Live Cell Imaging Solution

Image of cells stained with NucBlue Live reagent and propidium iodide
Jurkat cells stained with NucBlue® Live reagent and propidium iodide.

For Research Use Only. Not for use in diagnostic procedures.