The 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:

  • Cells growing in culture
  • One of the LIVE/DEAD® fixable dead cell stains (e.g. L34960)
  • Flow cytometer

Protocol tips

  • Cell concentration should be 1x104 to 1x106 cells per mL
  • Washing is required after staining
  • Cell staining is preserved after fixation, but fixation is not required
  • Dyes are provided in single-use vials and should be discarded after use

Labeling cells

 
1. Thaw vial of dye
 
2. Dilute LIVE/DEAD® fixable dead cell stain by adding 100 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 15 minutes
 

 

7. Wash cells

 


8. Analyze on flow cytometer

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 Dead Cell Stain Kit.


LIVE/DEAD® dye Excitation source Ex* Em*
Blue fluorescent UV 350 450
Violet fluorescent reactive dye 405 nm 416 451
Aqua fluorescent 405 nm 367 526
Yellow fluorescent 405 nm 400 575
Green fluorescent 488 nm 495 520
Red fluorescent 488 nm 595 615
Far red fluorescent 633/635 nm 650 665
Near-IR 633/635 nm 750 775
Sampler kit containing all 8 dyes see individual dye
* Approximate fluorescence excitation (Ex) and emission (Em) maxima, in nm.