Membrane integrity-based viability assay

The Invitrogen LIVE/DEAD fixable dead cell stains distinguish between live and dead cells in flow cytometry. The dyes covalently bind to intracellular and extracellular amines, allowing the staining pattern to be preserved following formaldehyde fixation. Choose from eight different fluorescent colors.

This protocol can be used for:

  • Identifying live and dead cells using a flow cytometer

This protocol should not be used for:

  • Fluorescence microscopy

You will need the following for this protocol:


1. Thaw vial of dye
2. Dilute LIVE/DEAD fixable dead cell stain by adding 50 µL DMSO to vial
3. Add 1 mL of cells to a flow cytometer tube in protein-free buffer
4. Add 1 µL of diluted stain to cells
5. Mix cells and stain
6. Incubate 30 minutes
7. Wash cells
8. Analyze on flow cytometer


Protocol tips

  • Cell concentration should be 1x104-1x106 cells per mL
  • Washing is optional after staining
  • Cell staining is preserved after fixation, but fixation is not required
  • Protein concentration in buffer should be <1%

Histogram showing live and dead cells distinguished using the LIVE/DEAD fixable dead cell stain kit reagents
Live and dead cells distinguished by flow cytometry using the LIVE/DEAD Fixable Violet Dead Cell Stain Kit.

Emission specifications

LIVE/DEAD dyeExcitation sourceEx*Em*
LIVE/DEAD Fixable Blue stainUV350450
LIVE/DEAD Fixable Violet stain405416451
LIVE/DEAD Fixable Lime stain405405506
LIVE/DEAD Fixable Aqua stain405367526
LIVE/DEAD Fixable Yellow stain405400575
LIVE/DEAD Fixable Green stain488495520
LIVE/DEAD Fixable Olive stain488480557
LIVE/DEAD Fixable Orange stain561580602
LIVE/DEAD Fixable Red stain488595615
LIVE/DEAD Fixable Far Red stain633/635650665
LIVE/DEAD Fixable Scarlet stain633/635700775
LIVE/DEAD Fixable Near-IR (775) stain633/635750775
LIVE/DEAD Fixable Near-IR (780) stain633/635750780
LIVE/DEAD Fixable Near IR (876) stain808840876
*Approximate fluorescence excitation (Ex) and emission (Em) maxima, in nm.