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Aurora cytometer

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The Aurora cytometer is a laboratory instrument designed for flow cytometry analysis. It is capable of detecting and analyzing cells, particles, and other biological samples. The Aurora cytometer provides accurate and reliable measurements of various cellular properties, such as size, granularity, and fluorescence intensity.

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Lab products found in correlation

Lysing solution, by BD (3 mentions) Celltrace cfse cell proliferation kit, by Thermo Fisher Scientific (3 mentions) Permeabilizing solution 2, by BD (3 mentions) Cytofix and phosflow perm wash buffers, by BD (3 mentions) Goat f ab 2 anti human iga g m, by Jackson ImmunoResearch (3 mentions) Alexa fluor 488 conjugated mab against phosphorylated, by BD (2 mentions) Alexa fluor 647 conjugated mab against phosphorylated, by BD (2 mentions) Leukocyte activation kit, by BD (1 mentions) Human tumor dissociation kit, by Miltenyi Biotec (1 mentions) Fc block reagent, by BD (1 mentions) Gentlemacs dissociator, by Miltenyi Biotec (1 mentions)

7 protocols using aurora cytometer

1

CFSE-Labeled PBMC Stimulation Assay

Cited 2 times
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PBMCs resuspended in PBS were labeled with 0.5 μM CFSE (CellTrace CFSE cell proliferation kit, ThermoFisher) at room temperature for 8 minutes, followed by addition of RPMI 1640–10% FBS and extensive washing as described (Jenks et al., 2018 (link); Kardava et al., 2018 (link)). The cells were then cultured with 2.5 μg/ml R848 and 1000 U/ml recombinant human IL-2 for 4 days at 1 × 106 cells per well of a 48-well flat-bottom plate. The cells were collected and stained with mAbs against CD19, CD20, CD3, CD27, CD21, IgD, CD38, and fluorochrome-conjugated spike proteins RBD-B.1 and S1-B.1, fixed (Lysing Solution, BD Biosciences), permeabilized (Permeabilizing Solution 2; BD Biosciences) and stained with mAbs against IgG, IgA, IgM. The cells were acquired on an Aurora cytometer and the effect of stimulation on cell division was evaluated by analysis using FlowJo v10.
Buckner C.M., Kardava L., Merhebi O.E., Narpala S.R., Serebryannyy L., Lin B.C., Wang W., Zhang X., de Assis F.L., Kelly S.E., Teng I.T., McCormack G.E., Praiss L.H., Seamon C.A., Rai M.A., Kalish H., Kwong P.D., Proschan M.A., McDermott A.B., Fauci A.S., Chun T.W, & Moir S. (2022). Recent SARS-CoV-2 infection abrogates antibody and B-cell responses to booster vaccination. medRxiv.
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2

CFSE-Labeling and Proliferation Assay

Cited 1 time
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PBMCs resuspended in PBS were labeled with 0.5 μM CFSE (CellTrace CFSE cell proliferation kit, ThermoFisher) at room temperature for 8 minutes, followed by addition of RPMI 1640–10% FBS and extensive washing as described (Kardava et al., 2018 (link)). The cells were then cultured with 2.5 μg/ml R848 and 1000 U/ml recombinant human IL-2 for 4 days at 1 × 106 cells per well of a 48-well flat-bottom plate. The cells were collected and stained with mAbs against CD19, CD20, CD3, CD27, CD21, IgD, CD38, and fluorochrome-conjugated spike proteins RBD-B.1 and S1-B.1, fixed (Lysing Solution, BD Biosciences), permeabilized (Permeabilizing Solution 2; BD Biosciences) and stained with mAbs against IgG, IgA, IgM. The cells were acquired on an Aurora cytometer and the effect of stimulation on cell division was evaluated by analysis using FlowJo v10. The division index, a measure of the overall proliferative response, is the average number of divisions undergone per cell in the total population, including cells that have not undergone division.
Buckner C.M., Kardava L., El Merhebi O., Narpala S.R., Serebryannyy L., Lin B.C., Wang W., Zhang X., Lopes de Assis F., Kelly S.E., Teng I.T., McCormack G.E., Praiss L.H., Seamon C.A., Rai M.A., Kalish H., Kwong P.D., Proschan M.A., McDermott A.B., Fauci A.S., Chun T.W, & Moir S. (2022). Interval between prior SARS-CoV-2 infection and booster vaccination impacts magnitude and quality of antibody and B cell responses. Cell, 185(23), 4333-4346.e14.
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3

Phosphorylation of Syk and PLC-γ2 in BCR-stimulated PBMCs

Cited 1 time
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PBMCs stained with mAbs against CD19, CD20, CD3, CD27, CD21, IgD, and BV421-conjugated RBD-B.1 were resuspended in RPMI 1640–10% FBS and stimulated with anti-BCR as described previously (Kardava et al., 2018 (link)), with the following modifications. The cells were stimulated at 37°C for 2 min with 10 μg/ml goat F(ab’)2 anti-human IgA/G/M (# 109-006-064 from Jackson ImmunoResearch Laboratories). For the detection of phosphorylated signalling intermediates, cells were fixed and permeabilized using BD Cytofix and Phosflow Perm/Wash buffers (BD Biosciences) and stained with Alexa Fluor 488-conjugated mAb against phosphorylated Syk (p-Y348) and Alexa Fluor 647-conjugated mAb against phosphorylated PLC-2 (p-Y759) (BD Biosciences). The samples were acquired on an Aurora cytometer and analysis was performed by FlowJo V10.
Buckner C.M., Kardava L., Merhebi O.E., Narpala S.R., Serebryannyy L., Lin B.C., Wang W., Zhang X., de Assis F.L., Kelly S.E., Teng I.T., McCormack G.E., Praiss L.H., Seamon C.A., Rai M.A., Kalish H., Kwong P.D., Proschan M.A., McDermott A.B., Fauci A.S., Chun T.W, & Moir S. (2022). Recent SARS-CoV-2 infection abrogates antibody and B-cell responses to booster vaccination. medRxiv.
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4

Phosphorylation of Syk and PLCγ2 in BCR-Stimulated PBMCs

Cited 1 time
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PBMCs stained with mAbs against CD19, CD20, CD3, CD27, CD21, IgD, and BV421-conjugated RBD-B.1 were resuspended in RPMI 1640-10% FBS and stimulated with anti-BCR as described previously (Kardava et al., 2018 (link)), with the following modifications. The cells were stimulated at 37°C for 2 min with 10 μg/ml goat F(ab’)2 anti-human IgA/G/M (# 109-006-064 from Jackson ImmunoResearch Laboratories). For the detection of phosphorylated signaling intermediates, cells were fixed and permeabilized using BD Cytofix and Phosflow Perm/Wash buffers (BD Biosciences) and stained with Alexa Fluor 488-conjugated mAb against phosphorylated Syk (p-Y348) and Alexa Fluor 647-conjugated mAb against phosphorylated PLCγ2 (p-Y759) (BD Biosciences). The samples were acquired on an Aurora cytometer and analysis was performed by FlowJo V10.
Buckner C.M., Kardava L., El Merhebi O., Narpala S.R., Serebryannyy L., Lin B.C., Wang W., Zhang X., Lopes de Assis F., Kelly S.E., Teng I.T., McCormack G.E., Praiss L.H., Seamon C.A., Rai M.A., Kalish H., Kwong P.D., Proschan M.A., McDermott A.B., Fauci A.S., Chun T.W, & Moir S. (2022). Interval between prior SARS-CoV-2 infection and booster vaccination impacts magnitude and quality of antibody and B cell responses. Cell, 185(23), 4333-4346.e14.
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5

Comprehensive Single-Cell Profiling of Tumor Immune Landscape

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Fresh tumor tissues were processed into single-cell suspensions using a gentleMACS™ Dissociator with a Human Tumor Dissociation Kit (Miltenyi Biotec). Two flow cytometry panels focused on T cells and myeloid cells were applied to single-cell suspensions of blood and tumors. All cell suspensions were initially incubated with a Leukocyte Activation Kit (BD Pharmingen) at 37 °C for 5 h and then divided into two tubes (1 × 106 cells/tube). Each tube was stained for viability and blocked with the Fc-block reagent (BD Pharmingen). The cells were then stained for surface markers (Additional file 1: Table S1-2). After fixation and permeabilization, cells were stained for intracellular markers (Additional file 1: Table S1-2). The limits for quadrant markers were always set based on negative populations. Cells were acquired using a Cytek Aurora cytometer and analyzed using FlowJo 10.8.0 software.
Han R., Guo H., Shi J., Zhao S., Jia Y., Liu X., Liu Y., Cheng L., Zhao C., Li X, & Zhou C. (2024). Osimertinib in combination with anti-angiogenesis therapy presents a promising option for osimertinib-resistant non-small cell lung cancer. BMC Medicine, 22, 174.
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6

CFSE-Labeling and Stimulation of PBMCs

Cited 1 time
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PBMCs resuspended in PBS were labeled with 0.5 μM CFSE (CellTrace CFSE cell proliferation kit, ThermoFisher) at room temperature for 8 minutes, followed by addition of RPMI 1640-10% FBS and extensive washing as described (Kardava et al., 2018 (link)). The cells were then cultured with 2.5 μg/ml R848 and 1000 U/ml recombinant human IL-2 for 4 days at 1 X 106 cells per well of a 48-well flat-bottom plate. The cells were collected and stained with mAbs against CD19, CD20, CD3, CD27, CD21, IgD, CD38, and fluorochrome-conjugated spike proteins RBD-B.1 and S1-B.1, fixed (Lysing Solution, BD Biosciences), permeabilized (Permeabilizing Solution 2; BD Biosciences) and stained with mAbs against IgG, IgA, IgM. The cells were acquired on an Aurora cytometer and the effect of stimulation on cell division was evaluated by analysis using FlowJo v10. The division index, a measure of the overall proliferative response, is the average number of divisions undergone per cell in the total population, including cells that have not undergone division.
Buckner C.M., Kardava L., El Merhebi O., Narpala S.R., Serebryannyy L., Lin B.C., Wang W., Zhang X., Lopes de Assis F., Kelly S.E., Teng I.T., McCormack G.E., Praiss L.H., Seamon C.A., Rai M.A., Kalish H., Kwong P.D., Proschan M.A., McDermott A.B., Fauci A.S., Chun T.W, & Moir S. (2022). Interval between prior SARS-CoV-2 infection and booster vaccination impacts magnitude and quality of antibody and B cell responses. Cell, 185(23), 4333-4346.e14.
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7

BCR Stimulation and Signaling Analysis

Cited 1 time
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PBMCs stained with mAbs against CD19, CD20, CD3, CD27, CD21, IgD, and BV421-conjugated RBD-B.1 were resuspended in RPMI 1640–10% FBS and stimulated with anti-BCR as described previously (Kardava et al., 2018 (link)), with the following modifications. The cells were stimulated at 37°C for 2 min with 10 μg/ml goat F(ab’)2 anti-human IgA/G/M (# 109-006-064 from Jackson ImmunoResearch Laboratories). For the detection of phosphorylated signaling intermediates, cells were fixed and permeabilized using BD Cytofix and Phosflow Perm/Wash buffers (BD Biosciences) and stained with Alexa Fluor 488-conjugated mAb against phosphorylated Syk (p-Y348) and Alexa Fluor 647-conjugated mAb against phosphorylated PLC-2 (p-Y759) (BD Biosciences). The samples were acquired on an Aurora cytometer and analysis was performed by FlowJo V10.
Buckner C.M., Kardava L., El Merhebi O., Narpala S.R., Serebryannyy L., Lin B.C., Wang W., Zhang X., Lopes de Assis F., Kelly S.E., Teng I.T., McCormack G.E., Praiss L.H., Seamon C.A., Rai M.A., Kalish H., Kwong P.D., Proschan M.A., McDermott A.B., Fauci A.S., Chun T.W, & Moir S. (2022). Interval between prior SARS-CoV-2 infection and booster vaccination impacts magnitude and quality of antibody and B cell responses. Cell, 185(23), 4333-4346.e14.
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