Clear Identification Of Dead Cells

April 04, 2018

Division of BioMedical Sciences, Faculty of Medicine
Memorial University of Newfoundland
Research Assistant

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Ease-of-Optimization

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Company:

BioLegend

Product Name:

Zombie Aqua Fixable Viability Kit

Catalog Number:

423101

Our lab routinely utilizes flow cytometry to investigate our research questions. Eliminating dead cells from flow cytometric data is necessary to obtain quality results, especially when working with in vitro stimulation assays or rare cell populations. The purpose of this experiment was to titrate the Zombie Aqua dye for optimal use to identify dead mouse splenocytes.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Live and heat-killed mouse splenocytes

Primary Incubation

30 min in the dark at room temperature with 1:100 dilution of Zombie Aqua dye diluted in PBS + 1mM EDTA (100ul total staining volume)

Blocking Agent

N/A

Secondary Incubation

N/A

Tertiary Incubation

N/A

Detection

Beckman Coulter MoFlow Astrios

Results Summary

Fresh and heat-killed mouse splenocytes were stained with 1:100 dilution of Zombie Aqua and analyzed after fixation and permeabilization (solid histograms). Cells alone without Zombie Aqua staining are indicated by the dashed histograms. In both cases, dead cells which stain positive could be easily distinguished from live cells. Staining was specific for dead cells since there was substantially more positively stained cells in the heat-killed splenocyte sample. 1:500 and 1:1000 dilutions also resulted in clear separation of live and dead cells.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Simple protocol; can be easilty integrated into existing cell staining protocols; can be titrated down to 1:500 or 1:1000 without compromising results

The Bad

None

The Bottom Line

Excellent product with little to no optimization required. Highly recommended.

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