LIVE/DEAD™ Fixable Near-IR Dead Cell Stain Kit, for 633 or 635 nm excitation

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Add the LIVE/DEAD Near‐IR stain reconstituted in DMSO to the cells at a final concentration of 1 : 10 000 Incubate for 30 min on ice and pellet by centrifugation (400 g, 10 min at +4 °C) and resuspend in 1 mL of PBS.	Detect at ~780 nm.

Protocol tips
Add the LIVE/DEAD Near‐IR stain reconstituted in DMSO to the cells at a final concentration of 1 : 10 000 Incubate for 30 min on ice and pellet by centrifugation (400 g, 10 min at +4 °C) and resuspend in 1 mL of PBS.
Downstream tips
Detect at ~780 nm.

Publication protocol

Dead cell discrimination (optional)A fixable dead cell stain can be added to the cell suspension. We used LIVE/DEAD Near‐IR (Invitrogen, Thermo Fischer Scientific, Waltham, MA, USA), which binds to amines on cell surface, but also intracellularly when the integrity of the cell is compromised. Intracellular amine staining yields an intense fluorescent signal in flow cytometry and allows the exclusion of dead cells from the downstream analyses.

2.1
Add the LIVE/DEAD Near‐IR stain reconstituted in DMSO to the cells at a final concentration of 1 : 10 000. Incubate for 30 min on ice and pellet by centrifugation (400 g, 10 min at +4 °C) and resuspend in 1 mL of PBS.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for LIVE/DEAD™ Fixable Near-IR Dead Cell Stain Kit, for 633 or 635 nm excitation below.

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