Accuri c6 plus flow cytometer

Manufactured by BD

Sourced in United States, Germany, United Kingdom, Japan, Italy

The BD Accuri C6 Plus flow cytometer is a compact, benchtop instrument designed for cell analysis. It utilizes flow cytometry technology to rapidly detect and analyze various cellular characteristics, including size, granularity, and fluorescence properties. The BD Accuri C6 Plus is capable of measuring multiple parameters simultaneously on individual cells within a sample.

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Lab products found in correlation

Accuri c6 plus software, by BD (32 mentions) Accuri c6, by BD (15 mentions) Propidium iodide, by Merck Group (13 mentions) Accuri c6 plus, by BD (11 mentions) Propidium iodide, by Thermo Fisher Scientific (9 mentions) Rnase a, by Merck Group (9 mentions) Trypsin edta, by Thermo Fisher Scientific (8 mentions) Annexin 5 fitc apoptosis detection kit, by Thermo Fisher Scientific (5 mentions) Goat anti mouse igg pe conjugated antibody, by BD (5 mentions) Facsdiva software, by BD (5 mentions) Annexin 5 fitc, by BD (4 mentions) Bovine serum albumin (bsa), by Merck Group (4 mentions) Binding buffer, by BD (4 mentions) Csampler plus software, by BD (4 mentions) Fitc annexin 5 apoptosis detection kit 1, by BD (4 mentions) Lipofectamine rnaimax, by Thermo Fisher Scientific (4 mentions) Sytox green, by Thermo Fisher Scientific (4 mentions) Rnase a, by Thermo Fisher Scientific (3 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Amaxa nucleofector device, by Lonza (3 mentions)

615 protocols using accuri c6 plus flow cytometer

Evaluating CD44-Mediated Uptake of HA-Complexes

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To determine the role of CD44 in the uptake of HA-formulated complexes, MDA-MB-231 was transfected with 60 nM CD44 siRNA at 1:6:1 siRNA:PEI-LA:HA w/w/w ratio. After 72 hrs, cells were exposed to FAM-labeled siRNA with 40 nM at 1:6:0 and 1:6:1 siRNA:PEI-LA:HA w/w/w ratio prepared as HA additive and coated complexes. The uptake of FAM-labeled siRNA was determined as described above using a BD Accuri™ C6 Plus Flow Cytometer (BD Biosciences, Franklin Lakes, NJ) after 4 hrs.
To determine the silencing efficiency of CD44 siRNA at the protein level, MDA-MB-231 cells were transfected with 60 nM CD44 siRNA at 1:6:0 and 1:6:1 siRNA:PEI-LA:HA w/w/w ratios. After 72 hrs, cells were washed with HBSS and incubated with PE-labeled anti-human control IgG and antihuman CD44 antibodies for 1 hr at room temperature. Cells were trypsinized and fixed with 3.7% formaldehyde. The surface binding of control IgG and CD44 antibodies were then quantified using BD Accuri™ C6 Plus Flow Cytometer (BD Biosciences).
The silencing efficiency of CD44 specific siRNA at the transcript level was determined by the Reverse Transcription -quantitative PCR (RT-qPCR). MDA-MB-231 cells were transfected with 60 nM CD44 siRNA at 1:6:0 and 1:6:1 siRNA:PEI-LA:HA w/w/w ratios. After 48 hrs, RT-qPCR was performed as described in RT-qPCR section below (Section 2.11).

Parmar M.B., Meenakshi Sundaram D.N., K C R.B., Maranchuk R., Montazeri Aliabadi H., Hugh J.C., Löbenberg R, & Uludağ H. (2018). Combinational siRNA delivery using hyaluronic acid modified amphiphilic polyplexes against cell cycle and phosphatase proteins to inhibit growth and migration of triple-negative breast cancer cells. Acta biomaterialia, 66.

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In Vitro Suppression and Proliferation Assay

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For the in vitro suppression assay, naïve T cells were treated with vehicle (DMSO) or a HDAC6 inhibitor (TSA), and then differentiated into iTreg cells for 3 days. Naïve T cells were stained with carboxy fluorescein succinimidyl ester (CFSE) (Sigma). Harvested iTreg and stained naïve T cells were cocultured in 96-well plates containing anti-CD3/CD28 beads (Invitrogen) (at several ratios). After 3 days, cells were harvested, and responder cells were selected and analyzed using a BD Accuri C6 Plus flow cytometer.
For the proliferation assay, naïve T cells were stained using CFSE (Sigma) and then polarized into each CD4 + T cell subset for 3 days. The stained cells were selected and analyzed using a BD Accuri C6 Plus flow cytometer.

Lee J.H., Kim H.S., Jang S.W, & Lee G.R. (2022). Histone deacetylase 6 plays an important role in TGF-β-induced murine Treg cell differentiation by regulating cell proliferation. Scientific Reports, 12, 22550.

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Evaluating TRPV1 and Mitochondrial Dysfunction in CML

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CML cells, treated with OLDA or vehicle at the IC₅₀ dose for 24 h, were incubated with 2 μg/mL PI for 30 min at 37°C. After washing, the fluorescence intensity was analyzed using BD Accuri C6 Plus software. The fluorescent probe DCFDA was used to assess oxidative stress levels in siTRPV1 and siGLO CML cells after treatment with OLDA (IC₅₀). Cells were incubated with 20 μM DCFDA for 20 min prior to the harvest time point. After washing, the fluorescence was assayed using the BD Accuri C6 Plus flow cytometer and its software. ∆Ψm was evaluated by JC-1 staining according to the manufacturer’s protocol in CML cells, treated with OLDA or vehicle at the IC₅₀ dose for 24 h. Samples were then analyzed using the BD Accuri C6 Plus flow cytometer and its software.

Maggi F., Morelli M.B., Aguzzi C., Zeppa L., Nabissi M., Polidori C., Santoni G, & Amantini C. (2023). Calcium influx, oxidative stress, and apoptosis induced by TRPV1 in chronic myeloid leukemia cells: Synergistic effects with imatinib. Frontiers in Molecular Biosciences, 10, 1129202.

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Cell Sorting of Xylanolytic Bacteria

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The cell sorting was realized from T. xylanilyticus cultivated on xylan basal medium at the generation 0 but also along generations (G20 and G50). The cell sorting was performed with a BD FACSAria™ II Cell Sorter coupled with the BD Accuri™ C6 Plus flow cytometer from the URCACyt technical platform facilities.
In order to standardize the signals of the BD Accuri™ C6 Plus flow cytometer and BD FACSAria™ II Cell Sorter, an analysis of 2.5 μm microbeads (BD Biosciences) was done with both systems.
5 × 10⁶ events were collected for each population and the rate of sorting was around 2500 events/s with a 70 μm nozzle. Each cell sorting was performed in triplicate. The events were collected in 10 mL of PBS 1×. To check for a correct cell sorting, an analysis of each population collected was done with BD Accuri™ C6 Plus flow cytometer. Each population solution was then centrifuged at 12,108×g (Sorvall ST 8R centrifuge, Thermo Scientific) for 30 min at room temperature. The cell pellet conserved in 100 µL of 1× PBS was solubilized with 1 mL of xylan basal medium. All the cell solution was used to inoculate a new cultivation on xylan and characterize the separated populations.

Bouchat R., Vélard F., Audonnet S., Rioult D., Delvigne F., Rémond C, & Rakotoarivonina H. (2022). Xylanase production by Thermobacillus xylanilyticus is impaired by population diversification but can be mitigated based on the management of cheating behavior. Microbial Cell Factories, 21, 39.

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Evaluating AML tumor burden in mice

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For HL-60-gfp-Luc þ orthotopic model, the bone, liver, and spleen were dissected at the termination of the experiment. Mononuclear cells collected from bone marrow (BM), spleen, and liver were analyzed by BD Accuri C6 Plus flow cytometer (BD Biosciences). The presence of GFP þ population was considered as AML tumor burden.
For PDX model, peripheral blood (PB) was weekly collected via retro-orbital bleeding. Bone, liver, and spleen were dissected at the termination of the experiment. Mononuclear cells collected from PB, BM, spleen, and liver were stained with FITC conjugated mouse anti-human CD45 (BioLegend). Stained cells were analyzed on the BD Accuri C6 Plus flow cytometer (BD Biosciences). The presence of the human CD45þ population was considered as AML tumor burden.

Cai Y., Chow J.P.H., Leung Y.O., Lu X., Yuen C.H., Lee W.L., Chau K.C., Yang L.L., Wong R.M.H., Lam J.Y.T., Chow D.T.L., Chung S.H.K., Kwok S.Y, & Leung Y.C. (2021). NEI-01-Induced Arginine Deprivation Has Potent Activity Against Acute Myeloid Leukemia Cells Both In Vitro and In Vivo. Molecular cancer therapeutics, 20(11).

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Quantitative Flow Cytometry Analysis

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The quantitative fluorescence of each cell line was estimated using a flow cytometer (BD Accuri^TM C6 Plus flow cytometer; BD Biosciences, USA) with BD Accuri^TM C6 Plus software (BD Biosciences, USA) and Flowjo software (BD Biosciences, USA). The cells were excited with a 488 nm laser and the emission was detected using a 533/30 nm bandpass filter (FITC-A). Details are provided in Supplementary Method 5. The gating strategy used for the analysis is described in Supplementary Fig. 9a–d.

Choi H.I., Hwang S.W., Kim J., Park B., Jin E., Choi I.G, & Sim S.J. (2021). Augmented CO2 tolerance by expressing a single H+-pump enables microalgal valorization of industrial flue gas. Nature Communications, 12, 6049.

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Cell Cycle and Apoptosis Analysis

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For cell-cycle analysis, HCT-8, HCT-8/5-FU, and DLD-1 cells were seeded in 6-cm dishes and cultured overnight, followed by treatment with vehicle control (DMSO) or a gradient concentration of CPX next day. After 24 h, cells were harvested and fixed in 70% ethanol and then stained with PI following RNase treatment. The stained cells were analyzed for cell-cycle distributions. For apoptosis assay, HCT-8, HCT-8/5-FU, and DLD-1 cells were incubated overnight, then treated with vehicle control (DMSO) or indicated concentration of CPX for 48 h. Cells were then collected and stained with Annexin V-FITC/PI for 20 min at room temperature.
Both cell-cycle distribution and cell apoptosis percentages were analyzed by flow cytometry on a BD Accuri^TM C6 plus flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA).

Qi J., Zhou N., Li L., Mo S., Zhou Y., Deng Y., Chen T., Shan C., Chen Q, & Lu B. (2020). Ciclopirox activates PERK-dependent endoplasmic reticulum stress to drive cell death in colorectal cancer. Cell Death & Disease, 11(7), 582.

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Apoptosis Evaluation by Flow Cytometry

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Both cells were treated with EPBS (0, 30, 50, and 100 µM) for 24 h. The cells were harvested and collected. The collected cells were stained with PI- and FITC-tagged annexin V antibodies for 15 min at RT. The annexin V/PI-stained cells were detected and analyzed using a BD Accuri^TM C6 Plus Flow Cytometer (BD Bioscience, Franklin Lakes, NJ, USA).

Kim N.Y., Mohan C.D., Chinnathambi A., Alharbi S.A., Sethi G., Rangappa K.S, & Ahn K.S. (2022). Euphorbiasteroid Abrogates EGFR and Wnt/β-Catenin Signaling in Non-Small-Cell Lung Cancer Cells to Impart Anticancer Activity. Molecules, 27(12), 3824.

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Apoptosis detection in breast cancer cells

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BT-474, SK-BR-3, MCF-7, and MDA-MB-231 cells were seeded on 6-well plates (1

\times

10⁶ cells/well) overnight and treated with 50 µM of PC-12 for 24 h. The cells were collected and fixed with 4% paraformaldehyde for 30 min at room temperature. After 30 min, the cells were washed with PBS and treated with 0.2% Triton x-100 for 10 min at RT. Then, the TUNEL enzyme and TUNEL label buffer were utilized for 1 h at 37 °C. The stained cells were detected and analyzed with a BD AccuriTM C6 Plus Flow Cytometer (BD Bioscience, Franklin Lakes, NJ, USA) [52 (link)].

Kim N.Y., Vishwanath D., Xi Z., Nagaraja O., Swamynayaka A., Kumar Harish K., Basappa S., Madegowda M., Pandey V., Sethi G., Lobie P.E., Ahn K.S, & Basappa B. (2023). Discovery of Pyrimidine- and Coumarin-Linked Hybrid Molecules as Inducers of JNK Phosphorylation through ROS Generation in Breast Cancer Cells. Molecules, 28(8), 3450.

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Cell Cycle Analysis of RPE-1 Cells

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150,000 RPE-1 cells were seeded into one well of a six-well plate for 18 h and treated with 25 ng/ml Mitomycin C (Sigma) or culture medium for 24 h. Cells were then harvested, washed once with PBS, and fixed in 70% EtOH. After two washes in 5 ml 1% BSA-PBS with a centrifugation at 1000 rpm 4 °C for 5 min, the cells were stained in PBS with 50 µg/ml propidium iodide (Sigma-Aldrich) and 0.1 µg/µl RNaseA (New England Biology) for 30 min at room temperature. At least 15,000 cells for each condition were analyzed in the BD Accuri^TM C6 Plus flow cytometer using the BD Accuri^TM C6 plus software.

Gao Y., Guitton-Sert L., Dessapt J., Coulombe Y., Rodrigue A., Milano L., Blondeau A., Larsen N.B., Duxin J.P., Hussein S., Fradet-Turcotte A, & Masson J.Y. (2023). A CRISPR-Cas9 screen identifies EXO1 as a formaldehyde resistance gene. Nature Communications, 14, 381.

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