Zombie nir fixable viability marker

Manufactured by BioLegend

The Zombie NIR Fixable Viability Marker is a fluorescent dye that can be used to detect dead cells in flow cytometry experiments. It binds to proteins in dead cells, allowing for the identification and exclusion of non-viable cells from analysis.

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Lab products found in correlation

Precision count beads, by BioLegend (2 mentions) Brilliant buffer, by BD (2 mentions) Spectroflo software, by Cytek (2 mentions) Cd8 bv421, by BioLegend (1 mentions) Aurora spectral flow cytometer, by Cytek (1 mentions) Cd14 percp cy5, by BioLegend (1 mentions) Cd19 apc, by BioLegend (1 mentions) Cd3 pe cy7, by BioLegend (1 mentions) Cd4 apc cy7, by BioLegend (1 mentions) Facs lysing solution, by BD (1 mentions) Cd45 fitc, by BioLegend (1 mentions)

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Zombie NIR (218 citations)

2 protocols using zombie nir fixable viability marker

Comprehensive Immune Cell Profiling in BAL

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Absolute cell counts in BAL were determined by flow cytometry with Precision Count Beads (BioLegend) by staining 100 μL of BAL fluid with pre-titrated amounts of CD45 FITC (clone HI30), CD14 PerCP-Cy5.5 (clone M5E2), CD3 PE-Cy7 (clone UCHT1), CD8 BV421 (clone RPA-T8), CD4 APC-Cy7 (clone OKT4) and CD19 APC (clone HIB19). 1 mL of FACS Lysing Solution (BD Biosciences) was added to each sample. 100 μL of Precision Count Beads were added immediately prior to flow cytometry to allow quantification of absolute volume analyzed on the cytometer.
In a separate experiment, fresh BAL cells were analyzed using a panel of antibodies directed against the following antigens: CD3 FITC (clone UCHT1), CD4 APC-Cy7 (clone OKT4), CD8 BV421 (clone RPA-T8), CD14 APC (clone M5E2), CD16 BV570 (clone 3G8), CD19 BV750 (clone HIB19), CD20 Pacific Blue (clone 2H7), CD38 PE-Cy7 (clone HIT2), CD45 Alexa Fluor 532 (clone HI30), CD56 PE/Dazzle 594 (clone HCD56), and HLA-DR BV605 (clone L243). 500,000–1,000,000 fresh BAL cells were stained in BD Brilliant Buffer (BD Biosciences) with Zombie NIR Fixable Viability Marker (BioLegend). Samples were run on a Cytek Aurora spectral flow cytometer using SpectroFlo software (version 2, Cytek) before final analysis in FlowJo software (version 10, BD Biosciences).

Reynolds D., Guillamet C.V., Day A., Borcherding N., Guillamet R.V., Choreño-Parra J.A., House S.L., O’Halloran J.A., Zúñiga J., Ellebedy A.H., Byers D.E, & Mudd P.A. (2021). Comprehensive immunologic evaluation of bronchoalveolar lavage samples from human patients with moderate and severe seasonal influenza and severe COVID-19. Journal of immunology (Baltimore, Md. : 1950), 207(5), 1229-1238.

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Multiparametric Immune Profiling of PBMCs

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Absolute counts of CD45⁺ cells in whole blood were determined at the time of blood collection on fresh samples by flow cytometry with Precision Count Beads (BioLegend). PBMCs, prepared using ficoll separation, were analyzed using a panel of antibodies directed against the following antigens: CD8 BV421 (clone RPA-T8), CD20 Pacific Blue (clone 2H7), CD16 BV570 (clone 3G8), HLA-DR BV605 (clone L243), immunoglobulin D (IgD) SuperBright 702 (clone IA6-2), CD19 BV750 (clone HIB19), CD45 Alexa Fluor 532 (clone HI30), CD71 PE (clone CY1G4), CD38 PE-Cy7 (clone HIT2), CD14 APC (clone M5E2), CD4 Spark 685 (clone SK3), and CD3 Alexa 700 (clone UCHT1). PBMC samples of 0.5 to 2 × 10⁶ cells were stained with a master-mix containing pretitrated concentrations of the antibodies, along with BD Brilliant Buffer (BD Biosciences) and Zombie NIR Fixable Viability Marker (BioLegend) to differentiate live and dead cells. Samples were run on a Cytek Aurora spectral flow cytometer using SpectroFlo software (Cytek) and unmixed before final analysis was completed using FlowJo software (BD Biosciences).

Mudd P.A., Crawford J.C., Turner J.S., Souquette A., Reynolds D., Bender D., Bosanquet J.P., Anand N.J., Striker D.A., Martin R.S., Boon A.C., House S.L., Remy K.E., Hotchkiss R.S., Presti R.M., O’Halloran J.A., Powderly W.G., Thomas P.G, & Ellebedy A.H. (2020). Distinct inflammatory profiles distinguish COVID-19 from influenza with limited contributions from cytokine storm. Science Advances, 6(50), eabe3024.

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