Image-iT DEAD Green Kit | Thermo Fisher Scientific

Membrane integrity-based viability assay

This kit includes Image-iT DEAD Green viability stain for identification of dead cells. After staining your cells, the signal from the stain will be retained if the sample is fixed and permeabilized.

This protocol can be used for:

Identifying dead cells using a fluorescence microscope

This protocol should not be used for:

Flow cytometry

You will need the following for this protocol:

Cells growing in culture
Image-iT DEAD Green Viability Stain (Cat. No. I10291)
Phosphate-buffered saline (PBS) (Cat. No. 14190-144)
Anhydrous DMSO (Cat. No. D12345)
Fluorescence microscope with FITC filters

Protocol

1. Culture cells in appropriate medium and vessel for microscopy

2. Dilute Image-iT DEAD Green stain by adding 225 µL DMSO to vial to make a 1,000X stock solution

3. Add 1 µL stock solution per milliliter of medium to label cells

4. Incubate 30 minutes at 37°C

5. Image cells

Spectral information and storage

	Image-iT DEAD Green
Excitation/Emission	488/515 nm
Standard filter set	FITC or GFP
EVOS Light Cube	GFP
Storage conditions	Room temperature

Protocol tips

This kit is compatible with antibody labeling protocols
Dye stock solutions should be stored frozen

Image of valinomycin-treated cells labeled with the Image-iT DEAD Green Kit reagents
Valinomycin-treated HeLa cells labeled with the Image-iT DEAD Green Viability Stain.

Image-iT DEAD Green Viability Stain Protocol