Two-color dual-parameter cell viability assay

This kit permits quick and easy determination of cell viability using two common microscope filters (FITC and TRITC) based on intracellular esterase activity and plasma membrane integrity.

This protocol can be used for:

  • Identifying live and dead cells using a fluorescence microscope

This protocol should not be used for:

  • Flow cytometry (see alternative flow cytometry protocol)

You will need the following for this protocol:


1. Culture cells in appropriate medium and vessel for microscopy
2. Thaw vials
3. Transfer Live Green vial (A) into Dead Red vial (B)
4. Mix to create 2X stock
5. Add equal volume 2X stock to cells
6. Incubate 15 min at 20–25°C
7. Image cells

Spectral information and storage

 Live GreenDead Red
Excitation/Emission488/515 nm570/602 nm
Standard filter setFITC or GFPTexas Red
EVOS Light CubeGFPTexas Red
Storage conditions–20°C–20°C


Protocol tips

  • Use 2x stock within 2 hours
  • Image quality may be improved by replacing media with Live Cell Imaging Solution
  • The stains do not survive fixation or permeabilization

Image of cells stained with LIVE/DEAD Cell Imaging Kit Reagents
Hep G2 cells stained using the LIVE/DEAD Cell Imaging Kit.