Accuri c6

Manufactured by BD

Sourced in United States, United Kingdom, Germany, China, France, Italy, Canada, Belgium, Japan, Switzerland, Australia, Ireland, Portugal, Spain, Uruguay, New Zealand

The BD Accuri C6 is a flow cytometer designed for analytical applications. It is capable of detecting and characterizing particles, cells, and other biological samples based on their size, granularity, and fluorescence properties.

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Lab products found in correlation

Accuri c6 flow cytometer, by BD (157 mentions) Propidium iodide, by Merck Group (94 mentions) Facscalibur, by BD (55 mentions) Fitc annexin 5 apoptosis detection kit, by BD (45 mentions) Annexin 5 fitc, by BD (41 mentions) Propidium iodide, by Thermo Fisher Scientific (39 mentions) Lsrfortessa, by BD (36 mentions) Rnase a, by Merck Group (35 mentions) Propidium iodide, by BD (32 mentions) Fitc annexin 5 apoptosis detection kit 1, by BD (30 mentions) Accuri c6 cytometer, by BD (28 mentions) Ionomycin, by Merck Group (28 mentions) Cflow plus software, by BD (27 mentions) Trypsin edta, by Thermo Fisher Scientific (24 mentions) Pi rnase staining buffer, by BD (23 mentions) 7 aad, by BD (22 mentions) Phorbol myristate acetate (pma), by Merck Group (22 mentions) Facsaria 3, by BD (22 mentions) Mitosox red, by Thermo Fisher Scientific (22 mentions) Brefeldin a, by Merck Group (21 mentions)

Spelling variants of this product

BD Accuri C6 Plus flow cytometer (615 citations) BD Accuri (258 citations) C6 Accuri software (9 citations) BD Accuri C6 1.0.264.21 (3 citations) BD Accuri C6 Sampler flow cytometer (3 citations) BD Accuri C6 FCM (2 citations) BD Accuri™ C6 CFlow Sampler flow cytometer (2 citations) Accuri C6 CFlow Sampler software (2 citations) Accuri C6 apparatus (2 citations) Accuri C6 ow cytometer and software (1 citations)

4 516 protocols using accuri c6

Flow Cytometry-based Cellular Assays

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All flow cytometry-based application were measured on a BD FACS Aria II or a BD Accuri C6 (BD Biosciences, Franklin Lakes, NJ, USA), per sample 10,000 cells were analyzed.
Annexin V/Propidium Iodide (PI) assay was performed as described previously [82 (link)]. Briefly, cells were trypsinized, pelleted and resuspended in FACS-buffer (10 mM HEPES/NaOH pH 7.4, 140 mM NaCl, 5 mM CaCl₂) containing Annexin V-APC (BD Biosciences) or Annexin V-Fluos (BD Biosciences) and PI and measured within 2 h.
EGFP-LC3-mRFP-LC3ΔG [54 (link)] fluorescence was measured on a BD Accuri C6 after the cells were harvested and resuspended PBS. The cells were measured in the FL-1 (EGFP) and FL-3 (mRFP) channels of a BD Accuri C6 and the ratio of EGFP/mRFP was calculated to estimate the autophagic flux.
Lysotracker^TM Deep Red (excitation/emission max 647/668 nM; Thermo Fisher) was used according to the manufacturer’s instruction. Briefly, 30 min prior to cell harvesting 25 nM LTDR was added and incubated at 37 °C in the dark. After trypsination, the cells were pelleted and washed twice with PBS and finally resuspended in 50 µL PBS for measurement in the FL-4 channel of a BD Accuri C6.

Remy J., Linder B., Weirauch U., Day B.W., Stringer B.W., Herold-Mende C., Aigner A., Krohn K, & Kögel D. (2022). STAT3 Enhances Sensitivity of Glioblastoma to Drug-Induced Autophagy-Dependent Cell Death. Cancers, 14(2), 339.

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Hemocyte Phagocytosis Assay by Flow Cytometry

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Hemolymph was extracted from the adductor muscle using 3 mL syringe with 23 G needle. The viability was determined by flow cytometry using a BD Accuri™ C6 (Becton Dickinson, San Jose, CA, USA) and by propidium iodide (PI) staining. Phagocytosis was evaluated according to the method developed by our laboratory.^{13 (link)} Briefly, hemocytes were mixed with yellow-green latex FluoSpheres (Molecular Probes Inc., Eugene, OR, USA) at a ratio of 1:100 (hemocytes:beads) in flat-bottom 96 wells plate. The mixtures were incubated at 20°C in the dark. After 18 h, the supernatant was delicately removed. The cells were fixed with 200 µL of 0.5% formalin in sterile water. Phagocytosis was measured by flow cytometry, BD Accuri™ C6 (Becton Dickinson) following the analysis of hemocytes according to their scattering properties of forward and right angle. A total of 3000 events were acquired to analyze fluorescence frequency distribution on FL1 and determined phagocytic activity (one bead and more) and phagocytic efficacy (three beads and more). Data collection and analysis were performed with BD Accuri™ C6 (Becton Dickinson) software.

Beaudry A., Lacaze E., Jobin-Piché A., Masson S., Auffret M., Brousseau P, & Fournier M. (2016). Ecotoxicological Evaluation of the Immunocompetence of two Bivalves Species (Mya Arenaria and Mytilus Edulis) in the Saguenay Fjord Including a Salinity Gradient. Journal of Xenobiotics, 5(2), 5769.

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Cell Sorting and Flow Cytometry

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Flow cytometry was performed on a BD Accuri C6 with manufacturer standard fluorescence detector set-up (FL1, 533/30 nm; FL2, 585/40 nm; FL3, > 670 nm; FL4, 675/25 nm) and data acquisition with BD Accuri C6 software including recording processed sample volume. Quality assurance was performed using fluorescent beads as per manufacturer protocol. Manual and extended cleaning cycles were performed at the beginning and end of each flow cytometry session with verification of low event rate in filtered PBS before each run. Cell-sorting experiments were performed on a Bio Rad S3 cell sorter or FACS Vantage with voltage and gain set to recreate BD Accuri C6 plots. All microscopy was performed on a Zeiss Axio Observer 7.

Barr D.A., Omollo C., Mason M., Koch A., Wilkinson R.J., Lalloo D.G., Meintjes G., Mizrahi V., Warner D.F, & Davies G. (2021). Flow cytometry method for absolute counting and single-cell phenotyping of mycobacteria. Scientific Reports, 11, 18661.

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Quantitative Analysis of Cell Viability

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For each extract, the rate of HAC/GFP loss was accurately measured independently using Synergy™ HTX Multi-Mode Microplate Reader (BioTek, United States) and BD Accuri C6 (BD Biosciences, United States) for a double check. For Synergy™ HTX Multi-Mode Microplate Reader (BioTek,United States), 1 × 10⁵ cells were seeded on a 96-well plate and depicted 24 h after extract treatment under 4х magnification in 5 × 5 fields of view. The number of GFP-positive and GFP-negative cells were counted in each field with threshold 7,000. The cell size was restricted from 5 μm up to 100 μm. For flowcytometry experiment using BD Accuri C6 (BD Biosciences, United States), 4 × 10⁵ cells per well were seeded on a 24-well plate with 500 μl of the culture medium and 10 μg/ml of Blasticidin S. Then cells were harvested by trypsin-treatment and resuspended in PBS containing 3 µM of DRUQ7. All the samples were vortexed immediately prior to flow cytometry examination using BD Accuri C6 (BD Biosciences, United States), the Fluorescence of GFP-positive cells was measured by the 488 nm laser and detected at 510 nm. The death cells were counted by DRUQ7 fluorescence excited by the 640 nm laser and detected at 722 nm. Samples were acquired in at least three separate triplicates for 30 s or 1.5 × 104 events (at minimum).

Goncharov N.V., Kovalskaia V.A., Romanishin A.O., Shved N.A., Belousov A.S., Tiasto V.S., Gulaia V.S., Neergheen V.S., Rummun N., Liskovykh M., Larionov V., Kouprina N, & Kumeiko V.V. (2022). Novel assay to measure chromosome instability identifies Punica granatum extract that elevates CIN and has a potential for tumor- suppressing therapies. Frontiers in Bioengineering and Biotechnology, 10, 989932.

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Flow Cytometry and Cell Sorting

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Flow cytometry was performed on a BD Accuri™ C6 with manufacturer standard fluorescence detector set-up (FL1, 533/30 nm; FL2, 585/40 nm; FL3, > 670 nm; FL4, 675/25 nm) and data acquisition with BD Accuri™ C6 software including recording processed sample volume. Quality assurance was performed using fluorescent beads as per manufacturer protocol. Manual and extended cleaning cycles were performed at the beginning and end of each flow cytometry session with verification of low event rate in filtered PBS before each run. Cell-sorting experiments were performed on a Bio Rad S3 cell sorter or FACS Vantage with voltage and gain set to recreate BD Accuri C6 plots. All microscopy was performed on a Zeiss Axio Observer 7.

Barr D., Omollo C., Mason M.K., Koch A., Wilkinson R.J., Lalloo D.G., Meintjes G., Mizrahi V., Warner D.F., & Davies G. (2021). Flow Cytometry Method for Absolute Counting and Single-Cell Phenotyping of Mycobacteria.

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Liposome-based pMHC Binding Assay

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Liposomes containing 1% DiO were formed with various ratios of DSPE-PEG2000-Maleimide (0, 0.1, 0.5, 1.0%) and reacted with Cys-terminated pMHC as described above. Naïve T cells (1×10⁶ cells/sample) were stained with 0–40 μg/ml pMHC liposomes in FACS buffer (1X DPBS + 2% FBS + 1mM EDTA + 25mM HEPES) for 30 min on ice. Cells were washed 4 times with 1 ml FACS buffer before analysis on a BD Accuri C6. Titration curves from median MFI were fit to one-site specific binding curve to determine the apparent dissociation constant (K_d,app).
For validation of antigen-specific binding, 1×10⁶ P14 or pmel splenocytes were co-stained with Db-GP33 liposomes (10 μg/ml) and anti-mouse CD8-PE (Biolegend, Clone: 53–6.7) for 30 min on ice. Cells were washed 3 times and analyzed on a BD Accuri C6. For competition experiments, P14 and pmel CD8⁺ cells were pre-stained with anti-mouse CD8-PE and anti-mouse CD8-APC, respectively, washed thoroughly with FACS buffer, and mixed together (1:1) in the same tube. The cells were stained with Db-GP33 liposomes as described above and analyzed on a BD Accuri C6. Gates were drawn around PE+ and APC+ cells and were used to quantify liposome uptake (DiO) in each group.

Dahotre S.N., Romanov A.M., Su F.Y, & Kwong G.A. (2021). Synthetic Antigen-Presenting Cells for Adoptive T Cell Therapy. Advanced therapeutics, 4(8), 2100034.

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Apoptosis and Cell Cycle Analysis

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For apoptosis assay, cells were stained with an Apoptosis Assay Kit (Wanlei Bio, Shenyang, China) according to the manufacturer’s instructions, and then detected by fluorescence-activated cell sorting (FACS) analysis. Briefly, cells were seeded in 6-well plates for lentivirus infection. Forty-eight hours later, cells were harvested and washed twice with cold phosphate buffered saline (PBS; double-helix, Shanghai, China). Subsequently, cells were incubated with 5 μl Annexin V-Light 650 and 10 μl propidium iodide (PI) in the dark for 15 min at room temperature. The apoptotic cells were detected by FACS analysis (Accuri C6, BD Biosciences) and the data were analyzed using BD Accuri C6 Software (BD Biosciences) on 10,000 events.
For cell cycle assay, cells were labeled with PI using a Cell Cycle Assay Kit (Beyotime) according to the manufacturer’s procedures. Briefly, 48 h after lentivirus infection, cells were harvested, washed twice with cold PBS, and fixed with ice-cold 70% ethanol for 2 h. Fixed cells were subsequently treated with 25 μl PI. Finally, 10 μl ribonuclease (RNase A) was added to the cells. The DNA content was then quantitated by FACS Accuri C6 (BD Biosciences), with an excitation wavelength of 488 nm and an emission wavelength of 625 nm. Data were analyzed using BD Accuri C6 Software (BD Biosciences) on 10,000 events.

Fu S., Fu Y., Chen F., Hu Y., Quan B, & Zhang J. (2018). Overexpression of MYCT1 Inhibits Proliferation and Induces Apoptosis in Human Acute Myeloid Leukemia HL-60 and KG-1a Cells in vitro and in vivo. Frontiers in Pharmacology, 9, 1045.

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Quantify CAR and FR Expression in SKOV3 Cells

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SKOV3 cells were seeded at a density of 1.5 × 10⁵ cells per well in a 96-well plate. Cells were washed in 200 µL of wash buffer (PBS/1% BSA) and incubated with 100 µL of wash buffer containing 1:500 of mouse anti-human monoclonal antibody against CAR (RmcB, Millipore, Watford, UK), 1:50 of mouse anti-human monoclonal anti-folate binding protein antibody (clone EPR4708(2)) (Abcam, UK) or mouse IgG control antibody (Santa Cruz Biotechnology, Heidelberg, Germany) for 1 h on ice. Cells were washed three times and incubated with a 1:500 dilution of goat anti-mouse Alexa Fluor 647 antibody (Invitrogen, UK) for 30 min on ice. Cells were fixed in 4% paraformaldehyde for 20 min at 4 °C. A total of 2 × 10⁴ gated events were acquired in channel FL-4 on a BD Accuri C6 (BD Biosciences, USA) flow cytometer and data analysed in BD Accuri C6 software version 1.0.264.21 (Becton Dickinson, USA). CAR and FR binding was analysed by flow cytometry relative to IgG isotype binding.

Hulin-Curtis S.L., Davies J.A., Nestić D., Bates E.A., Baker A.T., Cunliffe T.G., Majhen D., Chester J.D, & Parker A.L. (2020). Identification of folate receptor α (FRα) binding oligopeptides and their evaluation for targeted virotherapy applications. Cancer Gene Therapy, 27(10), 785-798.

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Viability Determination of Mytilus edulis Exposed to Lanthanides

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We know from some literature that lanthanides are not very cytotoxic. However, the viability was assessed in our experimental setting because it is a key element. Before doing any functional assay the viability must be determined. Measures were then taken on eight Mytilus edulis per lanthanide. Individuals were picked up at Mitis Bay (48°40’ N, 68°00’ W) and punctured blank with sterile 3 mL syringes and 2IG1½ needles. Cellularity and viability were measured by flow cytometry with BD Accuri™ C6 (Becton Dickinson, San Jose, CA, USA) and the software used was BD Accuri™ C6 Software (Becton Dickinson; version 1.0.264.21). Hemocyte populations were gated using a density plot (SSC vs FSC) and the fluorescence was recorded on FL3. Measurements were done using 200 L of cells exposed to propidium iodide at 10 µL*mL^–1 (Sigma-Aldrich Inc., St. Louis, MO, USA) and those samples with more than 70% of viable cells were processed further.

Pharand P., Brousseau-Fournier C., Côté C., Gagné F., Fournier M, & Brousseau P. (2016). Cellular Effects of Lanthanides on Mytilus Edulis. Journal of Xenobiotics, 5(2), 5781.

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TVRTSAEGGCGG Peptide Binding Assay

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SKOV3 were seeded at a density of 1 × 10⁵ cells per well in a 96-well plate. Cells were washed in 200 µL of wash buffer (PBS/1% BSA) and incubated with 100 µL of wash buffer containing either 100 µM, 300 µM or 500 µM TVRTSAEGGCGG peptide or PBS for 1 h at 4 °C. Cells were washed and incubated with wash buffer for 1 h at 4 °C. Cells were fixed in 4% paraformaldehyde for 20 min at 4 °C. A total of 2 × 10⁴ gated events were acquired in channel FL-4 on a BD Accuri C6 (BD Biosciences, USA) flow cytometer and data analysed in BD Accuri C6 software version 1.0.264.21 (Becton Dickinson, USA). TVRTSAEGGCGG peptide binding was analysed by flow cytometry relative to PBS control.

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