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Cflow plus software

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CFlow Plus software is a data analysis and visualization tool developed by BD for use with laboratory equipment. The software allows users to collect, analyze, and present data from various laboratory instruments.

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Lab products found in correlation

Accuri c6 flow cytometer, by BD (46 mentions) Accuri c6, by BD (27 mentions) Accuri c6 plus, by BD (6 mentions) C6 flow cytometer, by BD (4 mentions) Accuri c6 cytometer, by BD (4 mentions) Accuri flow cytometer, by BD (3 mentions) Fitc annexin 5 apoptosis detection kit 1, by BD (3 mentions) Anti ki67 fitc monoclonal antibody clone mib 1, by Agilent Technologies (2 mentions) Flourmount solution, by Merck Group (2 mentions) Annexin 5 fitc apoptosis detection kit, by Keygen Biotech (2 mentions) Accuri instrument, by BD (2 mentions) Propidium iodide solution, by Merck Group (2 mentions) Accuri c6 plus instrument, by BD (2 mentions) Apoptag fluorescein in situ apoptosis detection kit, by Merck Group (1 mentions) Annexin 5 kit, by BD (1 mentions) Annexin 5 staining kit, by BD (1 mentions) Centrifuge 5804 r, by Eppendorf (1 mentions) Lysing solution, by BD (1 mentions) Annexin 5 fitc kit, by BD (1 mentions) Stro 1 antibody, by R&D Systems (1 mentions)

Spelling variants of this product

CFlow software (71 citations) CFlow Plus (41 citations) CFlow (20 citations) CFlow Plus analysis software (11 citations) CFlow Plus program software (6 citations) Accuri CFlow Plus software (4 citations) BD Accuri CFlow Plus Analysis software (2 citations)

122 protocols using cflow plus software

1

Cisplatin-Induced Cell Cycle and Apoptosis

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For cell cycle distribution analysis Melanoma 7 cells (n.s., sh-catu2) were treated with cisplatin for 48 hours, detached, washed with PBS and fixed with 70% ice cold Ethanol. Then cells were washed again with PBS and stained with propidium-iodide solution (0.1% Triton-X-100, 2 mg DNAse free RNAse A and 500 μg/ml propidium-iodide) for 15 min at 37°C. Thereafter flow cytometry analysis was performed using Accuri Flow Cytometer and CFlow plus software (BD Biosciences).
For apoptosis-necrosis detection Melanoma 7 cells (n.s., sh-caut1 and 2) were treated with cisplatin for 48 h. Then cells were detached, centrifuged and washed with ice cold PBS. Thereafter cells were stained with APC-Annexin V (BD-Pharmingten) in 1x binding buffer (BD-Pharmingten) for 30 min at 4°C. Then Propidium Iodide Solution (Sigma-Aldrich) was added and Flow cytometry analysis was performed using Accuri Flow Cytometer and CFlow plus software (BD Biosciences).
Cytochrome c release assay was performed with the InnoCyte Flow Cytometric cytochrome c release Kit from Millipore (Billerica, MA, USA). Melanoma 7 cells (sh-catu2 and n.s.) were treated with different concentrations of cisplatin for 6 hours. Permeabilization, fixation and staining of the cells were performed according to the manufacturer’s instructions. Cells were analyzed using Accuri Flow Cytometer and CFlow plus software (BD Biosciences).
Kreiseder B., Holper-Schichl Y.M., Muellauer B., Jacobi N., Pretsch A., Schmid J.A., de Martin R., Hundsberger H., Eger A, & Wiesner C. (2015). Alpha-Catulin Contributes to Drug-Resistance of Melanoma by Activating NF-κB and AP-1. PLoS ONE, 10(3), e0119402.
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2

Cell Cycle and Apoptosis Analysis of 4-DACL

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For cell cycle distribution analysis NHM, A375 and IGR37 cells were starved for 24 h before cells were treated with 0, 1, 2.5 and 5µg/ml 4-DACL or left untreated for 24 hours. Cells were detached, washed with PBS and fixed with 70 % ice cold ethanol. Then cells were washed again with PBS and stained with propidium iodide solution (0.1 % Triton-X-100, 2 mg DNAse free RNAse A and 500 µg/ml propidium iodide) for 15 min at 37 °C. Thereafter flow cytometry analysis was performed using Accuri Flow Cytometer and CFlow plus software (BD Biosciences).
For apoptosis-necrosis detection IGR37 and A375 cells were treated with 1, 2.5, 5 and 10µg/ml 4-DACL or left untreated for 24 h. Then cells were detached, centrifuged and washed with ice cold PBS. Thereafter, cells were stained with APC-Annexin V (BD-Pharmingten) in 1x binding buffer (BD-Pharmingen) for 30 min at 4 °C. Then propidium iodide solution (Sigma-Aldrich) was added and flow cytometry analysis was performed using Accuri Flow Cytometer and CFlow plus software (BD Biosciences).
Genov M., Kreiseder B., Nagl M., Drucker E., Wiederstein M., Muellauer B., Krebs J., Grohmann T., Pretsch D., Baumann K., Bacher M., Pretsch A, & Wiesner C. (2016). Tetrahydroanthraquinone Derivative (±)-4-Deoxyaustrocortilutein Induces Cell Cycle Arrest and Apoptosis in Melanoma Cells via Upregulation of p21 and p53 and Downregulation of NF-kappaB. Journal of Cancer, 7(5), 555-568.
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3

Spleen Cell Characterization by Flow Cytometry

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Spleens were isolated, ground, and then filtered through a nylon sieve. Filtered cells were centrifuged at 1 000 g for 5 min. Single cells were suspended in red blood cell (RBC) lysis buffer to remove RBCs and were then cultured with RPMI 1640 complete medium for 16 h in the presence of protein transporter inhibitor. Cells were then harvested, washed twice with PBS and fixed with 4% PFA. Cell aliquots containing 5×105 cells were suspended in 100 μl of staining buffer, mixed with 1 μl of the corresponding fluorescent antibodies against surface markers, and then incubated on ice for 20 min. Cells were then washed with PBS twice and permeabilized for staining of intracellular markers. Flow cytometric analysis of cell surface markers included CD3, CD4, CD25 and intracellular molecules including IFN γ, Foxp3, IL-17. PE, FITC, or APC labeled CD3, CD4, CD25, IFN γ, Foxp3 and IL-17 antibodies, as well as corresponding isotype control antibodies, were all purchased from eBioscience (San Diego, CA). Samples were run in a flow cytometer (BD Accuri C6) and subsequently analyzed with BD CFlow Plus software.
Liu H., Zheng R., Wang P., Yang H., He X., Ji Q., Bai W., Chen H., Chen J., Peng W., Liu S., Liu Z, & Ge B. (2017). IL-37 Confers Protection against Mycobacterial Infection Involving Suppressing Inflammation and Modulating T Cell Activation. PLoS ONE, 12(1), e0169922.
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4

DNA Fragmentation Assay in MDA-MB 231 Cells

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To detect DNA fragmentation in MDA-MB 231 cells, a modified TUNEL method [37 ] was used with the ApopTag Fluorescein In Situ Apoptosis Detection Kit (Merck, KGaA, Darmstadt, Germany). Briefly, MDA-MB 231 cells were grown in 12-well culture plates and incubated with the compounds (10 or 30 μM) for 24 h. Following treatments, the cells were washed with PBS and subsequently stained with ApopTag according to the manufacturer’s instructions. Experiments were conducted in triplicate, 104 events/sample were analyzed by flow cytometry (BD Accuri C6 Plus, BD Biosciences, San Jose, CA, USA), and data were analyzed by BD CFlow Plus software (BD Biosciences, San Jose, CA, USA).
Guerra F.S., Dias F.R., Cunha A.C, & Fernandes P.D. (2021). Benzo[f]indole-4,9-dione Derivatives Effectively Inhibit the Growth of Triple-Negative Breast Cancer. Molecules, 26(15), 4414.
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5

Analyzing Cell Cycle Distribution by Flow Cytometry

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The cell cycle distribution was assessed using a BD Biosciences flow cytometry kit according to the manufacturer’s protocol (BD Biosciences, San Jose, CA, USA). Cells were seeded at 2.5 × 105 cells in 12-well plates and incubated with LASSBio-1911 or LASSBio-2208 at a final concentration of 0.1, 1, or 5 μM at 37 °C for 24 h in a humidified chamber containing 5% CO2. A total of 104 events of sample cells were recorded by flow cytometry (BD Accuri™ C6 Plus), and the data were analyzed by BD CFlow Plus software (BD Biosciences, San Jose, CA, USA).
Guerra F.S., Rodrigues D.A., Fraga C.A, & Fernandes P.D. (2021). Novel Single Inhibitor of HDAC6/8 and Dual Inhibitor of PI3K/HDAC6 as Potential Alternative Treatments for Prostate Cancer. Pharmaceuticals, 14(5), 387.
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6

Apoptosis Assay of LASSBio Compounds

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An apoptosis assay was carried out with an Annexin V kit (BD Biosciences, San Jose, CA, USA) by staining the cells according to the manufacturer’s protocol. Briefly, cells (2.5 × 105 cells) were seeded into 12-well plates and incubated with LASSBio-1911 or LASSBio-2208 at a concentration of 0.1, 1, or 5 μM for 24 h, harvested, washed with phosphate-buffered saline (PBS), and resuspended in the indicated binding buffer. Subsequently, Annexin V/FITC and propidium iodide (PI) were added to the cell suspension and incubated for 15 min in the dark at room temperature. After incubation, the samples were analyzed by flow cytometry (BD Accuri™ C6 Plus, BD Biosciences, San Jose, CA, USA), and the data were analyzed by BD CFlow Plus software (BD Biosciences, San Jose, CA, USA).
Guerra F.S., Rodrigues D.A., Fraga C.A, & Fernandes P.D. (2021). Novel Single Inhibitor of HDAC6/8 and Dual Inhibitor of PI3K/HDAC6 as Potential Alternative Treatments for Prostate Cancer. Pharmaceuticals, 14(5), 387.
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7

Annexin V Apoptosis Assay

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Apoptosis was carried out by an annexin V staining kit (BD Biosciences, San Jose, CA, USA) according to the manufacturer’s protocol. Briefly, cells (2.5 × 105 cells/mL) were seeded into 12-well plates and incubated with the compounds (at 10 or 30 μM) for 24 h. After the incubation period, the cells were harvested and washed with phosphate-buffered saline (PBS) and resuspended in the indicated binding buffer. Subsequently, annexin V-FITC and propidium iodide (PI) were added to the cell suspension and incubated for 15 min in the dark at room temperature. After incubation, 104 events/sample were analyzed by flow cytometry (BD Accuri C6 Plus, BD Biosciences, San Jose, CA, USA), and the data were analyzed by BD CFlow Plus software (BD Biosciences, San Jose, CA, USA).
Guerra F.S., Dias F.R., Cunha A.C, & Fernandes P.D. (2021). Benzo[f]indole-4,9-dione Derivatives Effectively Inhibit the Growth of Triple-Negative Breast Cancer. Molecules, 26(15), 4414.
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8

Multiparameter Flow Cytometry Protocol

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Cells were harvested and prepared for data collection as described for the small molecule screen. Centrifugation steps were conducted using two refrigerated table microcentrifuges: Preqlab Perfect Spin 24 R at 2000 rpm (376 rcf) and Eppendorf Centrifuge 5804 R at 1880 rpm (376 rcf) for 4 minutes. The stained cells were analyzed on a BD™ Accuri® C6 benchtop cytometer equipped with FL1 (533/30), FL2 (585/40) and FL4 (675/25) bandpass filters. Samples were run at 66 µL/min flow rate at 22 µm core size. Data were analyzed using BD™ CFlow® Plus software version 1.0.227.4 © 2008. Gates for detecting positive staining were set against unstained controls. A threshold of 0.5% of unstained cells within the positive gate was set. Where appropriate, compensation was applied according to single-stained control samples of the same cell type included in each individual experiment. The compensation values were generated to regain the set threshold of 0.5% in all channels (FL1, FL2, FL4). For very bright stainings, however, higher thresholds up to 1% within the positive gate were allowed after compensation. See Supplementary Table S3 for information on the antibodies used.
Ferlemann F.C., Menon V., Condurat A.L., Rößler J, & Pruszak J. (2017). Surface marker profiling of SH-SY5Y cells enables small molecule screens identifying BMP4 as a modulator of neuroblastoma differentiation. Scientific Reports, 7, 13612.
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9

Cell Cycle Analysis by Flow Cytometry

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The cell cycle was assessed using a BD Biosciences flow cytometry kit according to the manufacturer’s protocol (BD Biosciences, San Jose, CA, USA). Cells were seeded at 2.5 × 105 cells/well in 12-well plates and incubated with the compounds (10 or 30 μM) at 37 °C for 24 h in a humidified chamber containing 5% CO2. After staining the samples with PI (500 µg/mL) for 15 min, 104 events were analyzed by flow cytometry (BD Biosciences, San Jose, CA, USA), and the data were analyzed by BD CFlow Plus software (BD Biosciences, San Jose, CA, USA).
Guerra F.S., Dias F.R., Cunha A.C, & Fernandes P.D. (2021). Benzo[f]indole-4,9-dione Derivatives Effectively Inhibit the Growth of Triple-Negative Breast Cancer. Molecules, 26(15), 4414.
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10

Lymphocyte Flow Cytometry Assay

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Detached cells were resuspended in PBS. Lymphocytes were stained with conjugated, monoclonal antibodies against CD25 or CD69, respectively. The cells were assayed in a flow cytometre (Accuri™ C6, Becton Dickinson) and the data were analyzed with CFlow Plus software (Becton Dickinson).
Yin K., Zhu R., Wang S, & Zhao R.C. (2017). Low level laser (LLL) attenuate LPS-induced inflammatory responses in mesenchymal stem cells via the suppression of NF-κB signaling pathway in vitro. PLoS ONE, 12(6), e0179175.
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