C6 plus flow cytometer

Manufactured by BD

Sourced in United States

The C6 Plus flow cytometer is a compact and versatile instrument designed for cell analysis and sorting. It combines high-sensitivity detection with a user-friendly interface, providing researchers with accurate and reliable data. The C6 Plus is capable of analyzing a wide range of sample types, making it a valuable tool for various applications in the life sciences.

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Lab products found in correlation

Cellrox green reagent, by Thermo Fisher Scientific (1 mentions) Flow buffer, by Thermo Fisher Scientific (1 mentions) Flow buffer, by BD (1 mentions) Pe conjugated anti human cd19 antibody, by Thermo Fisher Scientific (1 mentions) Fluo 4, by Thermo Fisher Scientific (1 mentions) Fura red, by Thermo Fisher Scientific (1 mentions) Annexin 5 fitc binding solution, by Beyotime (1 mentions) Accuri c6 plus software, by BD (1 mentions) Accuri software, by BD (1 mentions) Annexin 5 fitc pi double staining kit, by Dojindo Laboratories (1 mentions) Coulter ac t diff 2, by Beckman Coulter (1 mentions) Cd42b apc, by BD (1 mentions) Pe annexin 5 apoptosis detection kit, by BD (1 mentions) Fixation permeabilization kit, by BD (1 mentions) Dcfh da, by Beyotime (1 mentions)

13 protocols using c6 plus flow cytometer

Evaluating Mhp390 Role in M. hyopneumoniae Adherence

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The role of mhp390 in M. hyopneumoniae adherence was evaluated through flow cytometry. PAM cells were seeded in 24-well plates, and then rMhp390 was incubated with PAM cells at 37°C for 30 min. Each well was washed for 5 times to remove the unbound protein. The bound rMhp390 was labeled green through anti His-tag mouse mAb and goat anti-mouse IgG-FITC. M. hyopneumoniae adhesin P97R1 and BSA served as the positive and negative control, respectively. The fluorescence intensity of PAM cells was evaluated with flow cytometry (BD C6 plus flow cytometer).
For the rMhp390 inhibition assay, M. hyopneumoniae was pre-blocked with two-fold serial dilution of rMhp390, which corresponded to 1.25–10.0 μg/ml of protein. BSA (10 μg/mL) and 1 mL of RPMI 1640 alone were used as the negative and blank controls, respectively. For the rMhp390 antiserum inhibition assay, M. hyopneumoniae was incubated with anti-rMhp390 serum diluted from 1:50 to 1:1000 before infection. The mixed negative serum (unimmunized rabbit serum, 1:50 dilution) was used as the negative control. The cells were evaluated with flow cytometry (BD C6 plus flow cytometer) and the adhesion ability of M. hyopneumoniae was evaluated based on the fluorescence intensity of PAM cells. These experiments were performed independently for three times.

Liu W., Jiang P., Yang K., Song Q., Yuan F., Liu Z., Gao T., Zhou D., Guo R., Li C., Sun P, & Tian Y. (2022). Mycoplasma hyopneumoniae Infection Activates the NOD1 Signaling Pathway to Modulate Inflammation. Frontiers in Cellular and Infection Microbiology, 12, 927840.

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Multiparameter Flow Cytometry Analysis

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Samples were fixed with 2% PFA for 20 min at RT, then wash with flow buffer (eBioscience). Samples were then incubated with PE-conjugated anti-human CD19 antibody (eBioscience) or isotype controls for 20 min at RT, then wash with flow buffer, followed by incubation with p62-Alexa Fluor 488 antibody for 60 min at RT. Samples were then washed with flow buffer, and analyzed with BD C6 plus flow cytometer.
For intracellular ROS measurement, 1X10⁶ cells in 500 µl medium per well were seeded in 24-well plates, and cultured overnight. 1 µl CellROX Green Reagent (Invitrogen) was added to each well and incubated for 30 min. Cells were then washed 3 times with PBS, and fixed with 2% PFA for 20min at RT, followed by extensive washes and then incubated with PE-conjugated anti-human CD19 antibody (eBioscience) for 20min at RT, before subjected to flow cytometry.

Wang L., Howell M.E., Sparks-Wallace A., Hawkins C., Nicksic C.A., Kohne C., Hall K.H., Moorman J.P., Yao Z.Q, & Ning S. (2019). p62-mediated Selective autophagy endows virus-transformed cells with insusceptibility to DNA damage under oxidative stress. PLoS Pathogens, 15(4), e1007541.

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Platelet Activation Assay by Flow Cytometry

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Whole blood was collected from anesthetized mice as previously described and diluted in modified Tyrode's buffer containing 1 mM CaCl 2 . Cell suspensions were then stimulated with ADP, MRS2365, Par4p or convulxin for 10 minutes at room temperature in the presence of 2 μg/mL PE-conjugated JON/A antibody (which detects activated αIIbβ3), 2.5 μg/mL of Alexa Fluor 488-conjugated anti-P-selectin antibody, and 2.5 μg/mL of Alexa Flour 647-conjugated anti-GPIX antibody. Samples were diluted with sterile-filtered PBS to stop the reaction and analyzed immediately by flow cytometry on a BD C6 Plus flow cytometer (BD Biosciences).

Paul D.S., Blatt T.N., Schug W.J., Clark E.G., Kawano T., Mackman N., Murcia S., Poe K.O., Mwiza J.M.N., Harden T.K., Bergmeier W, & Nicholas R.A. (2023). Loss of P2Y₁ receptor desensitization does not impact hemostasis or thrombosis despite increased platelet reactivity in vitro. Journal of thrombosis and haemostasis : JTH, 21(7).

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Platelets Calcium Mobilization Assay

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Washed platelets in Tyrode's buffer were diluted to 1 x 10 7 platelets/mL and loaded with 5 μM Fluo-4 and 5 μM Fura Red (Thermo Fisher Scientific, Waltham, MA) for 30 minutes at 37° C in the dark. Afterward, the samples were diluted to 1 x 10 6 platelets/mL in Tyrode's buffer and activated with the indicated concentrations of agonists in the presence of 1 mM Ca 2+ , while being continuously sampled by a BD C6 Plus flow cytometer. Kinetic calcium mobilization was analyzed in Flow Jo (Version 10) by plotting the ratio of Fluo4 to Fura Red mean fluorescence intensities (MFI) over time.

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Apoptosis Quantification by Flow Cytometry

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Cells were treated with Afzelin or transfected with NQO2 for 48 h and collected in a centrifuge tube. The cells were washed with PBS and digested with trypsin cell digestion solution. After centrifugation at 1000 g for 5 min, cells were collected, resuspended and counted to separate 5 × 10⁴ cells. The 5 × 10⁴ cells cells were centrifuged at 1000 g for 5 min and resuspended in Annexin V-FITC binding solution (Beyotime). Next, the cells were mixed with Annexin V-FITC and propidium iodide staining solution and incubated for 20 min at room temperature in the dark. After incubation, the apoptosis rate was measured using the BD C6 Plus flow cytometer (Becton, Dickinson and Company). The BD C6 Plus is a flow cytometry system that allows the measurement and analysis of various cellular parameters, including fluorescence signals emitted by labeled cells. Data acquisition and analysis was performed using BD Accuri C6 PLUS software.

Xia L., Xu X., Li M., Zhang X, & Cao F. (2023). Afzelin induces immunogenic cell death against lung cancer by targeting NQO2. BMC Complementary Medicine and Therapies, 23, 381.

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Annexin V-FITC/PI Apoptosis Assay

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After transfection with a plasmid, miRNA mimics, or siRNA for 48 h, the cells were collected and washed twice with phosphate-buffered saline at 4°C. The cells were then resuspended in binding buffer (1×), and adjusted to 1×10⁶ cells/mL. The cells were stained using an Annexin V-FITC/PI double staining kit (Dojindo, USA) and incubated in the dark for 15 min at room temperature. Apoptosis was analyzed by flow cytometry using a BD C6 Plus flow cytometer, and the percentage of cell apoptosis was analyzed using BD Accuri^TM software (both from Becton-Dickinson, CA, USA).

Qu L., Tian Y., Hong D., Wang F, & Li Z. (2022). Mig-6 Inhibits Autophagy in HCC Cell Lines by Modulating miR-193a-3p. International Journal of Medical Sciences, 19(2), 338-351.

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Platelet Activation and Reticulation

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Platelet count and mean platelet volume (MPV) were measured on a Coulter Ac T diff 2^® (Beckman Coulter, Brea, CA). Assessment of surface markers of platelet activation (P-selectin with and without exposure to thrombin, and CD40), and reticulated platelets (thiazole orange) was performed on a C6 Plus flow cytometer (BD Accuri, San Jose, CA) as previously described (10 (link)). Statistical comparisons were performed with independent samples t-tests (obese versus normal BMI) and paired-samples t-tests (obese before versus obese 6 months after surgery) using SPSS (Armonk, NY).

Heffron S.P., Marier C., Parikh M., Fisher E.A, & Berger J.S. (2018). Severe Obesity and Bariatric Surgery Alter the Platelet mRNA Profile. Platelets, 30(8), 967-974.

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Bariatric Surgery Patients' Platelet Activation

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Subjects were recruited from the Bellevue Hospital Center and NYU Langone Bariatric Surgery Clinics for an IRB-approved study. Non-diabetic, non-smoking individuals between 18 and 55 years of age otherwise meeting NIH criteria to undergo bariatric surgery (body mass index (BMI) ≥ 40kg/m², or ≥ 35kg/m² and at least one obesity-related comorbidity) were included [9 (link)]. Individuals on antiplatelet or anticoagulation medications, with NSAIDs use in the past 14 days, or on medications known to influence lipid levels were excluded.
Fasting blood samples were collected via a 19G needle at the initial preoperative visit. The first 2ml were discarded, the tourniquet removed, and the remaining blood was collected into tubes containing 3.2% (0.105M) sodium citrate as anticoagulant. Whole blood samples were exposed to thrombin at a submaximal concentration of 0.025U/mL (Instrumentation Laboratory, Bedford, MA) for five minutes or left untreated. Platelets were identified by staining with CD42b-APC (BD Biosciences) and platelet activation determined by platelet surface expression of P-selectin (FITC-conjugated anti-CD62P; BD Biosciences) on a C6 Plus flow cytometer (BD Accuri, San Jose, CA). A modified version of the National Cancer Institute Diet History Questionnaire II was used to estimate dietary composition at preoperative visits prior to dietary counseling.

Zhang R., Moscona A., Myndzar K., Luttrell-Williams E., Vanegas S., Jay M.R., Calderon K., Berger J.S, & Heffron S.P. (2021). More frequent olive oil intake is associated with reduced platelet activation in obesity. Nutrition, metabolism, and cardiovascular diseases : NMCD, 31(12), 3322-3325.

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Cell Cycle and Apoptosis Analysis

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Cells, after reaching 80% confluency, were washed twice with chilled PBS. To assess the cell cycle distribution, cell fixation was performed overnight at 4°C with pre-chilled 70% ethanol, followed by two chilled PBS washes and filtration with a 0.05 mm cell strainer. Next, the specimens were incubated with 50 μg/mL propidium iodide (PI), 100 μg/mL RNase A, and 0.2% Triton X-100 in PBS at 4°C for 30 min. Cellular DNA content was analyzed via flow cytometry on a C6 Plus flow cytometer (BD Biosciences, USA). To assess apoptosis, cells treated with EDTA-free trypsin were collected and stained with the PE Annexin V Apoptosis Detection Kit (BD Pharmingen) for 20 min at room temperature, as directed by the manufacturer.

Guo Z., Liang E., Li W., Jiang L, & Zhi F. (2021). Essential meiotic structure-specific endonuclease1 (EME1) promotes malignant features in gastric cancer cells via the Akt/GSK3B/CCND1 pathway. Bioengineered, 12(2), 9869-9884.

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Immunophenotyping of Activated Macrophages

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BMDMs were collected after LPS stimulation and treatment with PBS, SEVs, or HSEVs, as described above. Then the cells were permeabilized and fixed using a fixation/permeabilization kit (BD Bioscience, Cat# 554714) according to the manufacturer’s protocol. The cells were then incubated with rabbit anti-Arg1 antibody at 4°C for 30 minutes, washed with PBS, then incubated with PE donkey anti-rabbit antibody (1:200, eBioscience, Cat# 12-4739-81, RRID: AB_1210761) at 4°C for 30 minutes. After washing with PBS, the cells were incubated with CoraLite® plus 488 rabbit anti-iNOS antibody (1:500, Proteintech, Cat# CL488-18985, RRID: AB_2919161) and Alexa 647 anti-Iba1 antibody at 4°C for 30 minutes. Flow cytometry was performed using a C6 Plus flow cytometer (BD Biosciences), and data were analyzed using FlowJo software (TreeStar, Ashland, OR, USA).

Liang Z., Yang Z., Xie H., Rao J., Xu X., Lin Y., Wang C, & Chen C. (2024). Small extracellular vesicles from hypoxia-preconditioned bone marrow mesenchymal stem cells attenuate spinal cord injury via miR-146a-5p-mediated regulation of macrophage polarization. Neural Regeneration Research, 19(10), 2259-2269.

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