Cflow sampler software

Manufactured by BD

Sourced in United States

The CFlow Sampler software is a data acquisition and analysis tool for managing flow cytometry data. It provides functionalities for sample management, data visualization, and basic data analysis.

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Lab products found in correlation

Accuri c6 flow cytometer, by BD (6 mentions) Anti gata3 pe, by Thermo Fisher Scientific (4 mentions) Accuri c6, by BD (3 mentions) Anti cd4 percp, by Thermo Fisher Scientific (3 mentions) Anti tbet apc, by Thermo Fisher Scientific (3 mentions) Anti cd3 cd28 antibody, by Thermo Fisher Scientific (3 mentions) Accuri, by BD (3 mentions) Nylon cell strainer, by BD (3 mentions) Anti cd3 fitc, by Thermo Fisher Scientific (2 mentions) Penicillin, by Lonza (2 mentions) Streptomycin, by Lonza (2 mentions) Anti cd4 fitc, by Thermo Fisher Scientific (2 mentions) Anti mouse cd16 cd32, by Thermo Fisher Scientific (2 mentions) Red blood cell lysing buffer, by Merck Group (2 mentions) Automacs running buffer, by Miltenyi Biotec (2 mentions) Pd 1 antibody, by Cell Signaling Technology (1 mentions) Fxcycle pi rnase a staining solution, by Thermo Fisher Scientific (1 mentions) Cellinsight high content microscope, by Thermo Fisher Scientific (1 mentions) Click it edu alexa fluor 647 imaging kit, by Thermo Fisher Scientific (1 mentions) Alexa fluor plus 647, by Thermo Fisher Scientific (1 mentions)

16 protocols using cflow sampler software

Multicolor Flow Cytometry of Erythroid Precursors

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In the Indonesian cohort, pre-enriched (5 μL) and enriched (200,000 cells) suspensions from peripheral blood and sliced-spleen blood samples were stained with a 3-colour flow cytometry panel consisting of anti-CD45 (clone HI30) conjugated to Alexa Fluor 488, anti-CD71 (clone CY1G4) conjugated to phycoerythrin, and the fluorescent nucleic acid dye SYTO61. Isotype controls comprised anti-IgG2a conjugated to phycoerythrin instead of anti-CD71. Stains were incubated for 20 minutes at RT in the dark, then at least 150,000 events acquired on a BD Accuri C6 flow cytometer with CFlow Sampler software (BD Biosciences, Australia). All antibodies were purchased from BioLegend (San Diego, California) and SYTO61 from Thermofisher (Massachusetts, US).
In the French cohort, spleen fractions were centrifuged and 2 μL of each pellet resuspended in 1 mL of 1% Albumax-II/phosphate-buffered saline (PBS) (Thermofisher, Massachusetts, US). Suspensions were stained with anti-CD45 (clone HI30) conjugated to phycoerythrin-cyanine-7 and anti-CD71 (clone CY1G4) conjugated to allophycocyanin (both from BioLegend, San Diego, California) for 20 minutes at 4°C. Samples were washed, then resuspended in diluted BD Retic-Count (thiazole orange) and incubated in the dark for 1 hour at RT. At least 150,000 events were acquired on a BD Accuri C6 flow cytometer with CFlow Sampler software.

Kho S., Qotrunnada L., Leonardo L., Andries B., Wardani P.A., Fricot A., Henry B., Hardy D., Margyaningsih N.I., Apriyanti D., Puspitasari A.M., Prayoga P., Trianty L., Kenangalem E., Chretien F., Brousse V., Safeukui I., del Portillo H.A., Fernandez-Becerra C., Meibalan E., Marti M., Price R.N., Woodberry T., Ndour P.A., Russell B.M., Yeo T.W., Minigo G., Noviyanti R., Poespoprodjo J.R., Siregar N.C., Buffet P.A, & Anstey N.M. (2021). Evaluation of splenic accumulation and colocalization of immature reticulocytes and Plasmodium vivax in asymptomatic malaria: A prospective human splenectomy study. PLoS Medicine, 18(5), e1003632.

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FACS Analysis with BD Accuri C6

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FACS analysis was performed
in a BD Accuri C6 (San Jose, CA, USA) flow cytometer with CFlow Sampler
software (Becton Dickinson, Mountain View, USA).

Wu Y., Gu W., Chen C., Do S.T, & Xu Z.P. (2018). Optimization of Formulations Consisting of Layered Double Hydroxide Nanoparticles and Small Interfering RNA for Efficient Knockdown of the Target Gene. ACS Omega, 3(5), 4871-4877.

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PD-1 Expression in Mouse Lymphoma

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EL4 cells, which constitutively produce PD-1, a widely studied Mouse lymphoma cell line, was used as the cancer cell model in this study. Normally, EL4 cells were cultured in DMEM that was supplemented by 10% (v/v) FBS at 37 °C in 5% CO₂ atmosphere.
As-cultured EL4 cells were collected by centrifugation. After three washes with 2% FCS/PBS, the cells were fixed in 2% paraformaldehyde/PBS. For FACS analysis of PD-1 expression, the cells were then stained with PD-1 antibody (Cell Signal Technology, 1:500 dilution), and then analyzed in a BD Accuri™ C6 (San Jose, CA, USA) flow cytometer with CFlow Sampler software (Becton Dickinson, Mountain View, CA, USA).

Wu Y., Gu W., Li L., Chen C, & Xu Z.P. (2019). Enhancing PD-1 Gene Silence in T Lymphocytes by Comparing the Delivery Performance of Two Inorganic Nanoparticle Platforms. Nanomaterials, 9(2), 159.

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Flow Cytometric Analysis of Murine Lymphocytes

Cited 1 time

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We obtained mononuclear cells via the gentle extrusion of tissue from the spleen and the mesenteric lymph nodes (MLNs); the cells were analyzed using flow cytometry (Accuri, BD) and CFlow Sampler software (BD Biosciences) as described previously^{37 (link)}. Briefly, 1 × 10⁶–10⁷ cells were labeled with anti-CD3 FITC, anti-CD4 PerCP, anti-T-bet APC, and anti-GATA3-PE (all from eBioscience).
We performed stimulation experiments in which 2 × 10⁵ cells per well were stimulated with anti-CD3/CD28 antibodies (eBioscience, San Diego, USA) as described previously^{37 (link)}. We determined supernatant cytokine concentrations using a cytometric bead array system (Mouse Th1/Th2/Th17/Th22 13-Plex FlowCytomix Multiplex; eBioscience) in accordance with the manufacturer’s instructions.

Martín R., Chamignon C., Mhedbi-Hajri N., Chain F., Derrien M., Escribano-Vázquez U., Garault P., Cotillard A., Pham H.P., Chervaux C., Bermúdez-Humarán L.G., Smokvina T, & Langella P. (2019). The potential probiotic Lactobacillus rhamnosus CNCM I-3690 strain protects the intestinal barrier by stimulating both mucus production and cytoprotective response. Scientific Reports, 9, 5398.

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Flow Cytometry and Imaging Assays for Cell Proliferation

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Cells were fixed with 70% ethyl alcohol (EtOH) and stained with antibodies against ORF57 (sc-135746, SCBT) and anti-mouse IgG (H+L) Alexa Fluor Plus 647 (A-32728, Thermo Scientific), where indicated. DNA was then stained with FxCycle PI/RNaseA staining solution (Thermo Scientific) for 1 h. The cells were analyzed using a BD Accuri C6 flow cytometer and data were processed using the CFlow Sampler software (BD).
For cell proliferation analysis, cells were maintained for 2 h in medium containing 10 μM 5-ethynyl-2’-deoxyuridine (EdU). Subsequently, the cells were fixed in 4% paraformaldehyde in PBS and the EdU incorporated in the cell DNA was coupled to Alexa Fluor 647 according to the manufacturer’s instructions provided in the Click-iT EdU Alexa Fluor 647 imaging kit (Thermo Fisher Scientific). Images were acquired using a CellInsight High Content microscope (Thermo Fisher Scientific) and analyzed using Cell Profiler 3.0 software to quantify EdU-positive cells.

Elbasani E., Gramolelli S., Günther T., Gabaev I., Grundhoff A, & Ojala P.M. (2020). Kaposi’s Sarcoma-Associated Herpesvirus Lytic Replication Is Independent of Anaphase-Promoting Complex Activity. Journal of Virology, 94(13), e02079-19.

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Cell Cycle Analysis by PI Staining

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Cell cycle distribution was analyzed based on DNA staining using the Propidium Iodide (PI) Staining Solution (00-6990; eBioscience, San Diego, CA, USA) according to the manufacturer's protocol. Briefly, cells seeded in six-well plates were transfected with siRNAs and treated with bortezomib for 20 h. Cells were then washed with cold PBS, and fixed with cold 70% ethanol overnight. The PI staining was carried out by resuspending the cells in 500 μl PI/Triton X-100 solution (0.1% Triton X-100 in PBS, 0.2 mg/ml DNAse-free RNAse A, and PI Staining Solution diluted 1 : 200) for 30 min at room temperature. Cell cycle data were acquired using a BD Accuri C6 Flow Cytometer Instrument (BD Biosciences) and analyzed with the CFlow Sampler software (BD Biosciences).

Gu Y., Bouwman P., Greco D., Saarela J., Yadav B., Jonkers J, & Kuznetsov S.G. (2014). Suppression of BRCA1 sensitizes cells to proteasome inhibitors. Cell Death & Disease, 5(12), e1580-.

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Isolation and Flow Cytometric Analysis of Mesenteric Lymphoid Cells

Cited 1 time

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Mesenteric lymphoid nods (MLN) were gently extruded through a 50 μm-mesh Nylon cell strainer (BD) to isolate mononuclear cells. Then, cells were suspended in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% of FBS, 2 mM l-glutamine, 50 U/mg penicillin, and 50 µ/mg streptomycin (Lonza, Levallois-Perret, France). For flow cytometry analysis, aliquots of 10⁶–10⁷ cells per sample were labelled with anti-CD3-FICT, anti-CD4-PerCP, anti-Tbet-APC and anti-Gata3-PE according to the manufacturer’s instruction (eBioscience). The cells were analyzed using flow cytometry (Accuri, BD) and CFlow Sampler software (BD Biosciences) as described previously^{30 (link)}.

Martín R., Benítez-Cabello A., Kulakauskas S., Viana M.V., Chamignon C., Courtin P., Carbonne C., Chain F., Pham H.P., Derrien M., Bermúdez-Humarán L.G., Chapot-Chartier M.P., Smokvina T, & Langella P. (2023). Over-production of exopolysaccharide by Lacticaseibacillus rhamnosus CNCM I-3690 strain cutbacks its beneficial effect on the host. Scientific Reports, 13, 6114.

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Flow Cytometry Identification of DC and Treg Cells

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DC and Treg cells were identified by fresh whole blood flow cytometry using protocols detailed previously [14 (link)]. DC and Treg antibody panels are listed in Additional file 1. All samples were acquired on a portable BD Accuri C6 flow cytometer using CFlow Sampler Software (BD Biosciences). All antibodies were purchased from BioLegend (San Diego, CA, USA).

Kho S., Marfurt J., Handayuni I., Pava Z., Noviyanti R., Kusuma A., Piera K.A., Burdam F.H., Kenangalem E., Lampah D.A., Engwerda C.R., Poespoprodjo J.R., Price R.N., Anstey N.M., Minigo G, & Woodberry T. (2016). Characterization of blood dendritic and regulatory T cells in asymptomatic adults with sub-microscopic Plasmodium falciparum or Plasmodium vivax infection. Malaria Journal, 15, 328.

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Isolation and Characterization of Murine Immune Cells

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Mononuclear cells were isolated from spleens and MLN by gentle extrusion of the tissue through a 50 μm-mesh Nylon cell strainer (BD). Cells were suspended in Dulbecco’s Modified Eagle Medium (DMEM) medium supplemented with 10% of fetal calf serum (FCS), 2 mM L-glutamine, 50 U/mg penicillin, and 50 U/mg streptomycin (Lonza, Levallois-Perret, France). Erythrocytes were lysed with red blood-cell lysing buffer (Sigma–Aldrich).
For flow cytometry analysis, aliquots of 10⁶–10⁷ cells per sample were pre-incubated with purified anti-mouse CD16/CD32 (eBioscience, San Diego, CA, USA) and then labeled with anti-CD4-FITC, anti-CD3e-PE, and anti-CD8-PerCP (all from eBioscience) according to the manufacturer’s instructions. The stained cells were analyzed by flow cytometry (Accuri, BDbioscience) with CFlow Sampler software (BD).
For stimulation experiments, 2 × 10⁵ cells per well were cultured for 48 h (37°C, 10% CO₂) in DMEM medium in P24 plates pre-coated with anti-CD3/CD28 antibodies (4 μg/mL each; eBioscience) or phorbol 12-myristate 13-acetate (PMA)/ionomycin (cell stimulation cocktail, 1×, ebioscience). Culture supernatant was frozen at −80°C until processing.

Martín R., Laval L., Chain F., Miquel S., Natividad J., Cherbuy C., Sokol H., Verdu E.F., van Hylckama Vlieg J., Bermudez-Humaran L.G., Smokvina T, & Langella P. (2016). Bifidobacterium animalis ssp. lactis CNCM-I2494 Restores Gut Barrier Permeability in Chronically Low-Grade Inflamed Mice. Frontiers in Microbiology, 7, 608.

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Cell Cycle Analysis by Flow Cytometry

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Cells were harvested, fixed in 70% ethanol, stained with propidium iodide, and run on a BD Accuri C6 Flow Cytometer (BD Biosciences). DNA content from individual cells was analyzed using CFlow Sampler software (BD Biosciences), whereby the percentage of cells in G0/G1, G2/M, and apoptotic peaks was determined.

Gordon C.A., Gong X., Ganesh D, & Brooks J.D. (2017). NUSAP1 promotes invasion and metastasis of prostate cancer. Oncotarget, 8(18), 29935-29950.

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