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Attune nxt flow cytometer

Manufactured by BioLegend
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The Attune NxT Flow Cytometer is a versatile instrument designed for automated flow cytometry analysis. It offers high-speed data acquisition and advanced optics for sensitive and accurate detection of cells and particles.

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Lab products found in correlation

Human mouse th1 th2 th17 cytometric bead array, by BD (2 mentions) Ifn γ il 10 secretion assay detection kit, by Miltenyi Biotec (2 mentions) Facscanto 2, by BD (2 mentions) Legendplex human inflammation panel 1 13 plex, by BioLegend (2 mentions) Fcap array v3, by BD (2 mentions) Legendplex cloud based data analysis software, by BioLegend (2 mentions) Legendplex multiplex bead assay, by BioLegend (2 mentions) Legendplex data analysis software suite, by BioLegend (1 mentions) Custom mouse inflammation panel legendplex, by BioLegend (1 mentions) Legendplex mouse proinflammatory chemokine panel, by BioLegend (1 mentions) Legendplex data analysis suite, by BioLegend (1 mentions) Halt protease inhibitor, by Thermo Fisher Scientific (1 mentions)

6 protocols using attune nxt flow cytometer

1

Cytokine Profiling in Murine Inflammation

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Custom Mouse Inflammation Panel LEGENDplex (BioLegend, LEGENDplex™) protocol was followed according to the manufacturer's recommendations, and data was acquired using an Attune NxT Flow Cytometer. Pilot hind paw fluid samples from all experimental conditions were collected and used to determine sample dilution factors and to screen for 19 cytokines using capture beads targeting: GM-CSF, IL-1α, IL-1β, IL-4, IL-6, IL-9, IL-10, IL-12p70, IL-17A, IL-22, IL-23, IL-33, TNF-α, IFN-γ, CCL2, CXCL10, CCL4, CCL5, and LIF. We consistently detected only IL-1α in hind paw fluid in the capsaicin model. After initial screening, we selected 13 cytokines for ongoing analyses based on consistent detection in pilot studies and published literature indicating their potential role in neuroinflammation in skin. Biolegend LEGENDplex Data Analysis Software Suite (Qognit) was used to determine analyte mean fluorescence intensities and calculate concentrations based on concurrently-run standard curves (Supplementary File S3).
Salib A.M., Crane M.J., Lee S.H., Wainger B.J., Jamieson A.M, & Lipscombe D. (2024). Interleukin-1α links peripheral CaV2.2 channel activation to rapid adaptive increases in heat sensitivity in skin. Scientific Reports, 14, 9051.
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2

Cytokine Profiling by Flow Cytometry

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Supernatants from 96-well plates were aliquoted and stored at -20C. Cytokines were quantified by the Human/Mouse Th1/Th2/Th17 Cytometric Bead Array (CBA) (BD Biosciences) using a FACSCanto II (BD Biosciences), analyzed by FCAP Array v3.0 (BD Biosciences). Alternatively, we used the LEGENDplex Human Inflammation Panel 1 (13plex) (740808, BioLegend) with the Attune NxT Flow Cytometer. Cytokine assays analyzed by FlowJo/v9.
For intracellular cytokine detection, the IFN-γ/IL-10 Secretion Assay Detection Kit (Miltenyi, #130-090-761) was used on live cells after 72h.
Chauss D., Freiwald T., McGregor R., Yan B., Wang L., Nova-Lamperti E., Kumar D., Zhang Z., Teague H., West E.E., Vannella K.M., Ramos-Benitez M.J., Bibby J., Kelly A., Malik A., Freeman A.F., Schwartz D.M., Portilla D., Chertow D.S., John S., Lavender P., Kemper C., Lombardi G., Mehta N.N., Cooper N., Lionakis M.S., Laurence A., Kazemian M, & Afzali B. (2021). Autocrine Vitamin D-signaling switches off pro-inflammatory programs of Th1 cells. Nature immunology, 23(1), 62-74.
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3

Multiplex Cytokine Profiling of TLR3/MDA5 Activation

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We used a synthetic analog of dsRNA, poly(I:C), as a nonspecific agonist of TLR3 and MDA5/RIG-I. SV40-fibroblast cells were activated in 24-well plates, at a density of 100,000 cells/well, by incubation for 24 h with poly(I:C) at concentrations of 1, 5 and 25 μg/mL. Cells were stimulated with 25 μg/ml poly(I:C) in the presence of Lipofectamine 2000 to activate MDA5/RIG-I signaling. After 24 h, cell supernatants were used for the LEGENDplex multiplex bead assay (BioLegend, #740003 and #740984), then analyzed by flow cytometry on an Attune NxT Flow Cytometer, according to the manufacturer’s instructions. Data were analyzed with LEGENDplex Cloud-based Data Analysis Software (BioLegend), and presented in raw values and heatmaps. The heatmaps are generated using relative values for each sample as normalized to the maximum range of production levels of each cytokine among all samples (X-Min)/(Max-Min). Min: minimum production level; Max: maximum production level; X: the production level of a given cytokine in a given sample (96 ).
Liu Z., Garcia Reino E.J., Harschnitz O., Guo H., Chan Y.H., Khobrekar N., Hasek M.L., Dobbs K., Rinchai D., Materna M., Matuozzo D., Lee D., Bastard P., Chen J., Lee Y.S., Kim S.K., Zhao S., Amin P., Lorenzo L., Seeleuthner Y., Chevalier R., Mazzola L., Gay C., Stephan J.L., Milisavljevic B., Boucherit S., Rozenberg F., Perez de Diego R., Dix R.D., Marr N., Béziat V., Cobat A., Aubart M., Abel L., Chabrier S., Smith G.A., Notarangelo L.D., Mocarski E.S., Studer L., Casanova J.L, & Zhang S.Y. (2023). Encephalitis and poor neuronal death-mediated control of herpes simplex virus in human inherited RIPK3 deficiency. Science immunology, 8(82), eade2860.
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4

Lung Chemokine Profiling in Mice

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Lung tissue ∼50 mg was homogenized in PBS (500 μl) containing Halt protease inhibitor (Thermo Scientific) and centrifuged at 2000 rpm for 10 min. Lung homogenates (supernatant) were stored at −80°C until use. Chemokine analysis in lung homogenates was performed using the LEGENDplex mouse proinflammatory chemokine panel (BioLegend) according to the manufacturer’s protocol. Samples were run on Attune NxT flow cytometer and data were analyzed using the LEGENDplex data analysis suite (BioLegend).
Srinivasan A., Giri A., Duraisamy S.K., Alsup A., Castro M, & Sundar I.K. (2023). Chronic HDM exposure shows time-of-day and sex-based differences in inflammatory response associated with lung circadian clock disruption. iScience, 26(9), 107580.
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5

Cytokine Profiling by Flow Cytometry

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Supernatants from 96-well plates were aliquoted and stored at -20C. Cytokines were quantified by the Human/Mouse Th1/Th2/Th17 Cytometric Bead Array (CBA) (BD Biosciences) using a FACSCanto II (BD Biosciences), analyzed by FCAP Array v3.0 (BD Biosciences). Alternatively, we used the LEGENDplex Human Inflammation Panel 1 (13plex) (740808, BioLegend) with the Attune NxT Flow Cytometer. Cytokine assays analyzed by FlowJo/v9.
For intracellular cytokine detection, the IFN-γ/IL-10 Secretion Assay Detection Kit (Miltenyi, #130-090-761) was used on live cells after 72h.
Chauss D., Freiwald T., McGregor R., Yan B., Wang L., Nova-Lamperti E., Kumar D., Zhang Z., Teague H., West E.E., Vannella K.M., Ramos-Benitez M.J., Bibby J., Kelly A., Malik A., Freeman A.F., Schwartz D.M., Portilla D., Chertow D.S., John S., Lavender P., Kemper C., Lombardi G., Mehta N.N., Cooper N., Lionakis M.S., Laurence A., Kazemian M, & Afzali B. (2021). Autocrine Vitamin D-signaling switches off pro-inflammatory programs of Th1 cells. Nature immunology, 23(1), 62-74.
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6

Multiplex Cytokine Profiling of TLR3/MDA5 Activation

Check if the same lab product or an alternative is used in the 5 most similar protocols
We used a synthetic analog of dsRNA, poly(I:C), as a nonspecific agonist of TLR3 and MDA5/RIG-I. SV40-fibroblast cells were activated in 24-well plates, at a density of 100,000 cells/well, by incubation for 24 h with poly(I:C) at concentrations of 1, 5 and 25 μg/mL. Cells were stimulated with 25 μg/ml poly(I:C) in the presence of Lipofectamine 2000 to activate MDA5/RIG-I signaling. After 24 h, cell supernatants were used for the LEGENDplex multiplex bead assay (BioLegend, #740003 and #740984), then analyzed by flow cytometry on an Attune NxT Flow Cytometer, according to the manufacturer’s instructions. Data were analyzed with LEGENDplex Cloud-based Data Analysis Software (BioLegend), and presented in raw values and heatmaps. The heatmaps are generated using relative values for each sample as normalized to the maximum range of production levels of each cytokine among all samples (X-Min)/(Max-Min). Min: minimum production level; Max: maximum production level; X: the production level of a given cytokine in a given sample (96 ).
Liu Z., Garcia Reino E.J., Harschnitz O., Guo H., Chan Y.H., Khobrekar N., Hasek M.L., Dobbs K., Rinchai D., Materna M., Matuozzo D., Lee D., Bastard P., Chen J., Lee Y.S., Kim S.K., Zhao S., Amin P., Lorenzo L., Seeleuthner Y., Chevalier R., Mazzola L., Gay C., Stephan J.L., Milisavljevic B., Boucherit S., Rozenberg F., Perez de Diego R., Dix R.D., Marr N., Béziat V., Cobat A., Aubart M., Abel L., Chabrier S., Smith G.A., Notarangelo L.D., Mocarski E.S., Studer L., Casanova J.L, & Zhang S.Y. (2023). Encephalitis and poor neuronal death-mediated control of herpes simplex virus in human inherited RIPK3 deficiency. Science immunology, 8(82), eade2860.
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