Decent Fixable Viability Dye for Flow Cytometry

April 20, 2021

Emory University
Pediatrics
Graduate Student

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Company:

Biolegend

Product Name:

Zombie aqua fixable viability kit

Catalog Number:

423101/423102

Human PBMCs were isolated from blood, cultured in various conditions and stained for analysis by flow cytometry

Experimental Design and Results Summary

Application

Flow cytometry

Starting Material

Human PBMCs

Tips

be mindful of spillover into adjacent violet laser channels

Results Summary

Human PBMCs were isolated and cultured with differing stimuli and test compounds to investigate suppression of oxidative burst. Cells were harvested, washed, and stained for analysis of surface markers and intracellular targets by flow cytometry. The Zombie Live Dead Aqua dye was used to exclude dead cells from analysis. Live cells will appear as negative or weakly positive for this dye, while dead cells will be much higher in fluorescence due to compromised cell membrane

DOI or PMID #

N/A

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Summary

The Good

This stain accurately discriminates live and dead cells, and can be used with almost any cell type

The Bad

This is a degree of autofluorescence with myeloid cells in particular (monocytes and neutrophils), but this is common for many violet colors. Consider optimizing voltage settings for best results

The Bottom Line

This dye is reliable for excluding dead cells from flow cytometry analysis. However, the Zombie NIR may perform better for innate immune cells due to lower autofluorescence on occasion

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