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:


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

Spectral information and storage

 NucBlue LivePropidium iodide
Excitation/Emission360/460 nm535/617 nm
Standard filter setDAPITRITC or RFP
Storage conditionsRoom temperatureRoom 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
Jurkat cells stained with NucBlue Live reagent and propidium iodide.