Cyan adp flow cytometer

Manufactured by Beckman Coulter

Sourced in United States, Switzerland, United Kingdom, Italy, Australia, Canada

The CyAn ADP flow cytometer is a high-performance instrument designed for multiparameter analysis of cells and particles. It utilizes advanced fluidics and optics to provide precise and reliable measurements of fluorescence and light scatter properties.

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Lab products found in correlation

Zeba desalting column, by Thermo Fisher Scientific (7 mentions) Maleimide activated apc, by Abcam (7 mentions) Summit software v4, by Beckman Coulter (6 mentions) Propidium iodide, by Merck Group (5 mentions) Summit 4, by Beckman Coulter (5 mentions) Kaluza software, by Beckman Coulter (5 mentions) Summit version 4, by Beckman Coulter (5 mentions) Summit software, by Beckman Coulter (4 mentions) Sodium azide, by Merck Group (4 mentions) Summit v4, by Beckman Coulter (4 mentions) Bovine serum albumin (bsa), by Merck Group (4 mentions) Trypsin edta, by Thermo Fisher Scientific (4 mentions) Epcam, by Thermo Fisher Scientific (3 mentions) 7 aminoactinomycin d 7 aad, by Merck Group (3 mentions) Fcs5 express software, by De Novo Software (3 mentions) Cd11b, by Thermo Fisher Scientific (3 mentions) α sma, by Abcam (2 mentions) Epcam, by Abcam (2 mentions) Anti glast pe, by Miltenyi Biotec (2 mentions) Anti cd11b fitc, by BD (2 mentions)

Close spelling variants of this product

CyAn ADP (316 citations) CyAn flow cytometer (103 citations) CyAn ADP cytometer (33 citations) Cyan cytometer (13 citations) CyAn ADP Analyzer flow cytometer (11 citations) CyAn ADP High-Performance Flow Cytometer (5 citations) Cyan ADP 9-color flow cytometer (5 citations) Cyan ADP High Performance Research Flow Cytometer (4 citations) CyAn ADP 9C flow cytometer (3 citations) CyAn ADP Lx 9 color flow cytometer (3 citations)

214 protocols using cyan adp flow cytometer

Evaluating Cell Cycle and Cell Death

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For cell cycle analysis, cells were treated with OA at a concentration of 300 μM for 48 h and then the cells were fixed with 70% ethanol, washed three times with PBS and stained for 3 h at room temperature with PBS containing 20 μg/mL RNase A and 50 μg/mL propidium iodide (PI). Around 10,000 cells were analyzed using a CyAn ADP flow cytometer (Beckman Coulter, Brea, CA, USA) and FCS express 5 (De Novo software, Glendale, CA, USA). The experiment was performed three times with consistent results.
Annexin Pacific Blue /PI kit (Termo Fisher Scientific, Rockford, IL, USA) was employed for the detection of percentage of cell death according to manufacturer's instructions. Cells were treated with OA at the different concentrations into a 6-well plate at the density of 1 × 10⁵ cells/well for 24 h. Double staining was used to identify the cell membrane phosphatidylserine externalization and PI uptake. The results are from three independent experiments (n = 3). Samples were run on the CyAn ADP flow cytometer (Beckman Coulter) and analyzed with FlowJo software, version 10.5.3.

Giulitti F., Petrungaro S., Mandatori S., Tomaipitinca L., de Franchis V., D'Amore A., Filippini A., Gaudio E., Ziparo E, & Giampietri C. (2021). Anti-tumor Effect of Oleic Acid in Hepatocellular Carcinoma Cell Lines via Autophagy Reduction. Frontiers in Cell and Developmental Biology, 9, 629182.

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Quantifying GalNAc-T6 Expression on Cells

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Cells were fixed-permeabilized (4% PFA, 1% FBS, 0.1% Tween 20) and incubated with anti-GalNAc-T6 (T6.3) monoclonal antibody. The specific binding of primary MAb T6.3 to the cell lines was developed with an anti-mouse polyclonal antibody FITC-conjugated (Sigma-Aldrich; Merck KGaA) and further analyzed using a CyAnTM ADP Flow Cytometer (Beckman Coulter, Inc., Brea, CA, USA) and Summit v4.3 software. For each analysis 10,000 counts, gated on a FSC vs SSC dot plot excluding doublets populations, were recorded. Results were expressed as percentage of FITC positive cells and FITC mean fluorescence intensity (MFI). Data were expressed as the mean +/-standard deviation. Statistical analysis was determined using one-way analysis of variance (ANOVA) and consequently the Tukey's Multiple Camparison test using GraphPad Prism Sofware v5.00 Demo (GraphPad Software, Inc., La Jolla, CA, USA).

Ubillos L., Berriel E., Mazal D., Victoria S., Barrios E., Osinaga E, & Berois N. (2018). Polypeptide-GalNAc-T6 expression predicts better overall survival in patients with colon cancer. Oncology Letters, 16(1), 225-234.

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Quantification of IL-10-Producing CLL Cells

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Interleukin (IL)-10-competent CLL cell counts were determined by flow cytometry analysis of IL-10 production. Peripheral blood mononuclear cells (PBMCs) were purified from peripheral blood samples of the Dense-FCR arm of the protocol using Ficoll-Hypaque density gradients (Eurobio, Courtaboeuf, France).^{33 (link)} Clonal CLL cells were identified as CD19⁺ CD5⁺ CD20^int lymphocytes with a previously described protocol.^{34 (link)} Analyses were performed on a CyAnTM ADP flow cytometer (Beckman Coulter, Brea, CA, USA).

Gagez A.L., Duroux-Richard I., Leprêtre S., Orsini-Piocelle F., Letestu R., De Guibert S., Tuaillon E., Leblond V., Khalifa O., Gouilleux-Gruart V., Banos A., Tournilhac O., Dupuis J., Jorgensen C., Cartron G, & Apparailly F. (2017). miR-125b and miR-532-3p predict the efficiency of rituximab-mediated lymphodepletion in chronic lymphocytic leukemia patients. A French Innovative Leukemia Organization study. Haematologica, 102(4), 746-754.

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Intracellular ROS Generation in Heterophils

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To examine intracellular ROS generation, 1 × 10⁵ heterophils were seeded into
duplicate 96-well flat tissue culture plates. The cells were stimulated with PBS, 50
µM QH, 100 nM PMA, or 1 × 10⁶ cfu EC for 30
min (41°C, 5% CO₂). Following incubation, 10 µM
H₂DCF-DA was added to each well to stain the intracellular
H₂O₂ [7 (link), 8 (link)]. H₂DCF-DA readily diffuses into cells,
where it is oxidized to the highly fluorescent 2′,7′-dichlorofluorescein (DCF) [2 (link)]. Cells were incubated in the dark for another 5 min,
then washed with cold PBS, and sample acquisition (10,000 events) was performed on
ROS-containing cells using a CyAn^TM ADP Flow Cytometer (Beckman Coulter, Brea,
CA, U.S.A.).

NAMBOOPPHA B., PHOTICHAI K., WONGSAWAN K, & CHUAMMITRI P. (2018). Quercetin manipulates the expression of genes involved in the reactive oxygen species (ROS) process in chicken heterophils. The Journal of Veterinary Medical Science, 80(8), 1204-1211.

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Spleen Leucocyte Isolation and Analysis

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The spleen tissues were pressed through sterilized nylon mesh (40 µm, Dutscher)
with Leibovitz 15 (L15) medium (Sigma) containing heparin lithium (10 U.mL -1 , Sigma), penicillin (500 U.mL -1 , Biochrom AG) and streptomycin (500 µg.mL -1 , Biochrom AG) to obtain leucocyte suspension. After storage of 12 hours at 4 ± 1°C, L15 medium-diluted samples were loaded onto Ficoll gradient (Histopaque®1077, density of 1,077g.mL -1 , Sigma). After centrifugation (400 g, 30 min, 15 °C), leucocytes enriched suspensions were collected and washed twice in L15 medium (300 g, 5 min, 4 °C). Then, leucocytes were adjusted at 10 6 cells.mL -1 with Malassez haemocytometer to perform cytometer analyses.
Analyses were carried out on whole leucocytes, using a Cyan TM ADP flow cytometer (Beckman Coulter). For each leucocyte sample, 10,000 cells were counted.
Leucocyte differential was obtained using forward scatter (FSC) and size scatter (SSC) parameters for size and granularity, respectively. The cellular mortality percentage and the phagocytosis activity were determined, using propidium iodure (1.0 g.L -1 , Molecular Probes) and Fluorescent microsphere (2.7*10 10 particles.mL -1 , Fluorospheress carboxylatemodified microsphere, diameter 1 mm, Molecular Probes), respectively, according to Bado-Nilles et al. (2009a) .

Bado-Nilles A., Jolly S., Lamand F., Geffard A., Gagnaire B., Turies C., Porcher J.M., Sanchez W, & Betoulle S. (2015). Involvement of fish immunomarkers in environmental biomonitoring approach: Urban and agri-viticultural context. Ecotoxicology and environmental safety, 120.

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Apoptosis Assay with Annexin V-FITC

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Apoptosis was assessed with the Annexin V-FITC kit according to the manufacturer's instructions (BioVision, Milpitas, CA, USA). Dual staining with fluorescent Annexin V and propidium iodide (PI) has been used to discriminate apoptotic and necrotic cell death, in which Annexin V-positive/PI-negative staining is regarded as apoptosis and PI-positive staining as necrosis Cells were seeded on 25 cm 2 culture plates with 6.2.10 6 THP-1 cells, 0.8.10 6 A549 cells and 2.5.10 6 HPMEC-ST1.6R cells in 5 ml media per plate. After a 24 h stabilization, cells were exposed to a freshly prepared TiO 2 -NPs suspension in cell media (5, 200 and 800 µg/ml concentrations, corresponding to 1, 40, 160 µg/cm 2 ) for 24 h. Cells were washed twice with cold PBS, collected, and resuspended in binding buffer. Annexin V-FITC and propidium iodide were added, and the cells were incubated for 10 min at room temperature in the dark. The cells were calculated with flow cytometry (CyanTM ADP flow cytometer, Beckman Coulter).

Hanot-Roy M., Tubeuf E., Guilbert A., Bado-Nilles A., Vigneron P., Trouiller B., Braun A, & Lacroix G. (2016). Oxidative stress pathways involved in cytotoxicity and genotoxicity of titanium dioxide (TiO2) nanoparticles on cells constitutive of alveolo-capillary barrier in vitro. Toxicology in vitro : an international journal published in association with BIBRA, 33.

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Immune Cell Profiling in Cancer

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Flow cytometric analysis of immune cell populations and CAF was performed by labelling fluorescent cells with tagged antibodies against: CD45, CD4, CD8, CD19, NK1.1, CD11b, F4/80, Gr1, CD11c, MCH-II, EpCam, GP38 (eBioscience),, CD31 (BD, Franklin Lakes, NJ, USA), and α-SMA (Abcam), CAF were determined as CD45⁻EpCam⁻CD31⁻GP38⁺αSMA⁺ cells. Intracellular staining for RORγt and IL-17A, was performed on MLN and spleen single cell suspensions. Samples were analyzed on a Beckman Coulter Cyan ADP flow cytometer (Beckman Coulter, Indianapolis IN, USA), and data were analyzed with FlowJo.

Sanchez-Lopez E., Flashner-Abramson E., Shalapour S., Zhong Z., Taniguchi K., Levitzki A, & Karin M. (2015). Targeting colorectal cancer via its microenvironment by inhibiting IGF-1 Receptor-insulin receptor substrate and STAT3 signaling. Oncogene, 35(20), 2634-2644.

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Immune Cell Profiling in Cancer

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Quantifying Glial Cell Populations

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The estimated percent of glia in mixed glial, microglial, and astrocyte cultures were determined by immunophenotying using direct labeling with anti-GLAST-PE (Miltenyi Biotec, San Diego, CA), anti-Cd11b-FITC (BD Biosciences), anti-CD11b-PE (Stemcell Technologies), and anti-GLAST-488 (Novus Biologicals, Littleton, CO) followed by flow cytometric analysis. Cells were counted using a Bio-Rad TC10 automated cell counter, and 1 × 10⁶ cells/mL were resuspended in 100 μL of incubation buffer (PBS with 0.05% bovine serum albumin). Mixed glial cultures were labeled using the mouse anti-GLAST-PE (20 μg/mL) and mouse anti-Cd11b-FITC (10 μg/mL) at room temperature for 1 h. Microglia cultures were incubated with CD11b-PE according to manufacturer instructions while astrocyte cultures were incubated with rabbit polyclonal anti-GLAST-488 (10 μg/mL) at room temperature for 1 h. After labeling, the cells were washed twice in incubation buffer and resuspended at a final volume of 500 μL of PBS and stored at 37 °C until analysis. Flow cytometry was performed on a Beckman Coulter CyAn ADP flow cytometer operated with Summit software for data collection at Colorado State University’s Flow Cytometry Core Facility. All further data analysis was done utilizing FlowJo software (version 10.1; FlowJo, Ashland, OR).

Kirkley K.S., Popichak K.A., Afzali M.F., Legare M.E, & Tjalkens R.B. (2017). Microglia amplify inflammatory activation of astrocytes in manganese neurotoxicity. Journal of Neuroinflammation, 14, 99.

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Multiparametric Flow Cytometry Analysis

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Primary antibodies: anti-MET (Human HGFR/c-MET APC-conjugated Antibody, clone 95,106, R&D System); anti-IgG1/CH2CH3 regions (Alexa Fluor® 647 AffiniPure F (ab’)2 Fragment Goat Anti-Human IgG (H + L) antibody, Jackson Immuno Research); anti-CD4 (APC Mouse Anti-Human CD4 clone M-T466, Miltenyi); anti-CD3 (PE Mouse Anti-Human CD3, Clone HIT3a); anti-CD8 (APC Mouse Anti-Human CD8, Clone RPA-T8); anti-CD56 (PE Mouse Anti-Human CD56, Clone MY31) all from BD Biosciences. Isotype control antibodies: APC, FITC, or PE mouse IgG1 κ Isotype Control, Clone MOPC-21 (BD Biosciences). Cells were counterstained with DAPI and analyzed by Cyan ADP flow cytometer (Beckman Coulter S.r.l.). Data were elaborated using Summit 4.3 software (Dako). For plots in which the isotype control is not shown, the Mean Fluorescence Intensity (MFI) derived from the Isotype control was set within the first logarithm (0 < MFI < 10). Cells were considered positive for the analyzed marker if the signal was higher than 10 (MFI > 10).

Chiriaco C., Donini C., Cortese M., Ughetto S., Modica C., Martinelli I., Proment A., Vitali L., Fontani L., Casucci M., Comoglio P.M., Giordano S., Sangiolo D., Leuci V, & Vigna E. (2022). Efficacy of CAR-T immunotherapy in MET overexpressing tumors not eligible for anti-MET targeted therapy. Journal of Experimental & Clinical Cancer Research : CR, 41, 309.

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