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Facs accuri c6

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The FACS Accuri C6 is a flow cytometer designed for basic cell analysis and sorting. It is capable of detecting and quantifying various cellular parameters, such as size, granularity, and fluorescence intensity, using laser-based technology. The instrument is intended for use in research and clinical laboratory settings.

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Lab products found in correlation

Accuri c6, by BD (6 mentions) Cflow sampler v 1, by BD (4 mentions) Propidium iodide, by BD (3 mentions) Fitc annexin 5 apoptosis detection kit, by BD (3 mentions) Dcfh da, by Merck Group (2 mentions) Fcap array software, by BD (2 mentions) Cell cycle assay kit, by Beyotime (2 mentions) Anti pd l1, by Miltenyi Biotec (2 mentions) Annexin 5 fluos staining kit, by BD (2 mentions) Annexin 5 apoptosis detection kit, by Thermo Fisher Scientific (2 mentions) Annexin 5 fitc apoptosis detection kit, by Merck Group (1 mentions) Lyse fix solution, by BD (1 mentions) Accutase, by Merck Group (1 mentions) Fcr blocking reagent, by Miltenyi Biotec (1 mentions) Perm buffer 3, by BD (1 mentions) Sp8 falcon, by Leica (1 mentions) Annexin 5 fitc pi apoptosis kit, by Solarbio (1 mentions) Cell cycle kit, by Beyotime (1 mentions) Alexa fluor 647 conjugated anti active caspase 3 antibody, by BD (1 mentions) Anti mhcii fitc, by BD (1 mentions)

75 protocols using facs accuri c6

1

Quantifying IL-8 in HIV-1 Infection

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Plasma levels of IL-8 were measured using Cytometric Bead Flex Set (BD Biosciences, Sparks, MD, USA) in the HIV-1 infected children and seronegative donors. Briefly, 50 μL of beads coated with capture anti IL-8 antibodies (capture beads), were mixed with 50 μL of sample and incubated for 1 hour in dark at room temperature. After 1 hour, 50 μL of phycoerythrin conjugated detection antibody was added to this sample-bead mixture, incubated for 2 hours in the dark at room temperature. The samples were washed with 1 mL of wash buffer (300 g, 5 minutes) and were then acquired on the FACS Accuri C6 and analyzed using FCAP Array Software (BD Biosciences, Sparks, MD, USA). Calibration curves were constructed using serial dilutions of cytokine standard provided with the kit. Instrument calibration was done (BD FACS Accuri C6) with SPHERO 8-peak Rainbow Particles to ensure accuracy/validity of the data.
In order to evaluate the changes in the relative mRNA expression and plasma levels of IL-8 with time, we followed up 15 ART progressors post 6 months’ initiation of ART.
Pananghat A.N., Aggarwal H., Prakash S.S., Makhdoomi M.A., Singh R., Lodha R., Ali S., Srinivas M., Das B.K., Pandey R.M., Kabra S.K, & Luthra K. (2016). IL-8 Alterations in HIV-1 Infected Children With Disease Progression. Medicine, 95(21), e3734.
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2

Palmitate-induced Apoptosis in MIN6 Cells

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MIN6 cells were stimulated with palmitate (0.4 mM) for 36 h in the presence and absence of GRP75 siRNA or were transfected with the empty vector or GRP75 overexpression vector, and apoptosis was detected using the FITC Annexin V Apoptosis Detection Kit (Sigma) according to the manufacturer’s instructions. Another set of cells transfected with the scramble or the GRP75 siRNA (100 nM: 48 h) alone was similarly analyzed for the detection of apoptosis. Briefly, cells were harvested by trypsinization and cells was resuspended in the binding buffer containing annexin V-FITC and PI conjugate and incubated for 10 min at room temperature. Cells were then analyzed for apoptosis by flow cytometry (BD FACS Medoly and BD FACS Accuri C6, BD Biosciences). Data was acquired and normalized to equal number of cells.
Tiwary S., Nandwani A., Khan R, & Datta M. (2021). GRP75 mediates endoplasmic reticulum–mitochondria coupling during palmitate-induced pancreatic β-cell apoptosis. The Journal of Biological Chemistry, 297(6), 101368.
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3

Annexin V and PI Staining for Cell Death

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The cells were infected under a MOI of 10 and 1, prepared using supernatants from infected C6/36, and equal volume of mock. Cellular infection occurred at 4°C for 1 hour with cell homogenization every 10 minutes. After that, the cells were washed once and then maintained at 37°C in CO2 incubators with medium, as described before. After 24, 48, 72 and 96 hours p.i. the cells were harvested and then submitted to a staining protocol for annexin V and propidium iodide (PI) (BD Biosciences®). The cells were washed twice with PBS and were harvested with 200 μL of trypsin 0.25 % (LGC®) for 10 minutes at 37°C. Next, the cell suspensions were washed in DMEM with 10% of FBS and centrifuged for 5 minutes at 450 g and 4°C. The cells were then ressuspended in 20 μL of annexin V binding buffer in 96-well round bottom plates and with 1 μL of FITC-annexin V + 1 μL of PI and then incubated at room temperature for 15 minutes protected from light. After incubation period, the samples were added to 80 μL of binding buffer and acquired in the BD FACS Accuri C6 (BD Biosciences®) flow cytometer.
Cugola F.R., Fernandes I.R., Russo F.B., Freitas B.C., Dias J.L., Guimarães K.P., Benazzato C., Almeida N., Pignatari G.C., Romero S., Polonio C.M., Cunha I., Freitas C.L., Brandão W.N., Rossato C., Andrade D.G., Faria D.D., Garcez A.T., Buchpigel C.A., Braconi C.T., Mendes E., Sall A.A., Zanotto P.M., Peron J.P., Muotri A.R, & Beltrão-Braga P.C. (2016). The Brazilian Zika virus strain causes birth defects in experimental models. Nature, 534(7606), 267-271.
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4

Annexin V and PI Staining for Cell Death

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The cells were infected under a MOI of 10 and 1, prepared using supernatants from infected C6/36, and equal volume of mock. Cellular infection occurred at 4°C for 1 hour with cell homogenization every 10 minutes. After that, the cells were washed once and then maintained at 37°C in CO2 incubators with medium, as described before. After 24, 48, 72 and 96 hours p.i. the cells were harvested and then submitted to a staining protocol for annexin V and propidium iodide (PI) (BD Biosciences®). The cells were washed twice with PBS and were harvested with 200 μL of trypsin 0.25 % (LGC®) for 10 minutes at 37°C. Next, the cell suspensions were washed in DMEM with 10% of FBS and centrifuged for 5 minutes at 450 g and 4°C. The cells were then ressuspended in 20 μL of annexin V binding buffer in 96-well round bottom plates and with 1 μL of FITC-annexin V + 1 μL of PI and then incubated at room temperature for 15 minutes protected from light. After incubation period, the samples were added to 80 μL of binding buffer and acquired in the BD FACS Accuri C6 (BD Biosciences®) flow cytometer.
Cugola F.R., Fernandes I.R., Russo F.B., Freitas B.C., Dias J.L., Guimarães K.P., Benazzato C., Almeida N., Pignatari G.C., Romero S., Polonio C.M., Cunha I., Freitas C.L., Brandão W.N., Rossato C., Andrade D.G., Faria D.D., Garcez A.T., Buchpigel C.A., Braconi C.T., Mendes E., Sall A.A., Zanotto P.M., Peron J.P., Muotri A.R, & Beltrão-Braga P.C. (2016). The Brazilian Zika virus strain causes birth defects in experimental models. Nature, 534(7606), 267-271.
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5

Intracellular ROS Detection in HSC-3 Cells

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To monitor the generation of intracellular ROS, viable HSC-3 cells were pre-treated with NAC for 1 h at 37°C, followed by 10 and 20 µM of LH for 24 h. The generation of intracellular ROS was detected via flow cytometry with 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) as the peroxide-sensitive fluorescent probe, which may be converted to DCFH and then oxidized to the fluorescent compound DCF in the presence of ROS in cells. The cells treated with NAC and LH were harvested, washed with PBS, mixed with 10 µM DCFH-DA and incubated at 25°C for 30 min in the dark. The cell suspension was subjected to the flow cytometer and the fluorescence signal was detected for intracellular ROS measurement via flow cytometry (BD FACS Accuri C6; BD Biosciences) and data analysis of intracellular ROS was performed using a CFlow Sampler v.1.0 software (BD Biosciences).
Li M.H., Liao X., Li C., Wang T.T., Sun Y.S., Yang K., Jiang P.W., Shi S.T., Zhang W.X., Zhang K., Li C, & Yang P. (2021). Lycorine hydrochloride induces reactive oxygen species-mediated apoptosis via the mitochondrial apoptotic pathway and the JNK signaling pathway in the oral squamous cell carcinoma HSC-3 cell line. Oncology Letters, 21(3), 236.
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6

Apoptosis analysis of LH-treated HSC-3 cells

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Viable HSC-3 cells treated with different concentrations of LH (with or without pre-treatment of 5 mM NAC for 1 h) for 24 h were harvested and washed with ice-cold PBS. Cells were dual-stained with Annexin V-FITC and PI at 25°C for 20 min, followed by FCM analysis according to the manufacturer's protocol of the Annexin V-FITC/PI apoptosis detection kit. Data acquisition and analysis of the cell apoptosis were performed using a flow cytometer (BD FACS Accuri C6; BD Biosciences) and CFlow Sampler v.1.0 software (BD Biosciences).
Li M.H., Liao X., Li C., Wang T.T., Sun Y.S., Yang K., Jiang P.W., Shi S.T., Zhang W.X., Zhang K., Li C, & Yang P. (2021). Lycorine hydrochloride induces reactive oxygen species-mediated apoptosis via the mitochondrial apoptotic pathway and the JNK signaling pathway in the oral squamous cell carcinoma HSC-3 cell line. Oncology Letters, 21(3), 236.
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7

Measuring Mitochondrial Membrane Potential

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The changes in MMP were detected via staining cells with JC-1, a fluorescent probe for MMP detection. HSC-3 cells treated with LH (with or without pre-treatment of 5 mM NAC for 1 h) for 24 h were harvested, washed with ice-cold PBS and stained with 5 mg/ml JC-1 at 37°C for 30 min in the dark. Data acquisition and analysis of MMP were performed by flow cytometry (BD FACS Accuri C6; BD Biosciences) and CFlow Sampler v.1.0 software (BD Biosciences).
Li M.H., Liao X., Li C., Wang T.T., Sun Y.S., Yang K., Jiang P.W., Shi S.T., Zhang W.X., Zhang K., Li C, & Yang P. (2021). Lycorine hydrochloride induces reactive oxygen species-mediated apoptosis via the mitochondrial apoptotic pathway and the JNK signaling pathway in the oral squamous cell carcinoma HSC-3 cell line. Oncology Letters, 21(3), 236.
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8

Cell Cycle and Apoptosis Analysis

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To determine the distribution of the cells in the cell cycle and the proportion of apoptotic cells, we performed flow cytometry analysis using a flow cytometer (BD FACS Accuri C6, CA, USA). After a 24 h treatment with ATL (0, 10 and 20 μM), the cells were collected, washed with PBS and fixed with ice-cold 70% ethanol at 4 °C for 4 h. The cells were stained with propidium iodide (PI) staining buffer (0.2% Triton X-100, 100 μg/mL DNase-free RNase A, and 50 μg/mL PI in PBS) in the dark for 30 min. For the apoptosis examination, the cells were washed with PBS, collected, and stained using the Annexin V-FITC Apoptosis Detection Kit in the dark at room temperature for 15 min. The cell cycle distribution and the fraction of apoptotic cells were determined using a FACS analysis system. Each experimentwas performed in triplicate.
Wang X., Yu Z., Wang C., Cheng W., Tian X., Huo X., Wang Y., Sun C., Feng L., Xing J., Lan Y., Sun D., Hou Q., Zhang B., Ma X, & Zhang B. (2017). Alantolactone, a natural sesquiterpene lactone, has potent antitumor activity against glioblastoma by targeting IKKβ kinase activity and interrupting NF-κB/COX-2-mediated signaling cascades. Journal of Experimental & Clinical Cancer Research : CR, 36, 93.
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9

Intracellular ROS Detection in ZF4 Cells

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To monitor the intracellular generation of ROS, viable ZF4 cells were collected for staining. The generation of intracellular ROS was detected using 2′,7′-Dichlorodihydrofluorescein diacetate (DCFH-DA, D6883, Sigma, MO, United States) as the peroxide-sensitive fluorescent probe. The cell suspension (5 × 104 cells/mL) was incubated with 10 μM DCFH-DA for 30 min in the absence of any light. After being washed with 1 × PBS, the cell suspension was analyzed by flow cytometry within 30 min (BD FACS Accuri C6; BD Biosciences, CA, United States). Lastly, data analysis of intracellular ROS was conducted using FlowJo 10 software.
Wang H., Wang Y., Niu M., Hu L, & Chen L. (2022). Cold Acclimation for Enhancing the Cold Tolerance of Zebrafish Cells. Frontiers in Physiology, 12, 813451.
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10

Murine Bone Marrow-Derived Macrophage Immunophenotyping

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Adherent mouse BMDMs were detached with accutase (#A6964, Sigma) for 20 min at 37°C and blocked with FcR Blocking reagent (#130-092-575, Miltenyi Biotec) for 15 min at 4°C. Antibodies used in flow cytometry were anti-CD11b (M1/70), anti-CD11c (HL3), anti-CD86 (GL1), anti-CD40 (HM40-3) from BD Biosciences; anti-TLR4 (MTS510) from Biolegend; anti-IFNGR1 (2E2) from eBioscience; anti-iNOS (M-19) from SCT and anti-goat AlexaFluor 647 Donkey (polyclonal IgG H&L) from Abcam (#ab150131). For intracellular staining of iNOS, macrophages were fixed with BD Lyse/Fix solution (#558049) for 10 min at 37°C, then permeabilized with BD Perm Buffer III (#558050) for 20 min at 4°C prior to antibody staining. Flow cytometric acquisitions were performed on a BD FACS Accuri C6 and data were analyzed using FlowJo software.
Oldenburg R., Mayau V., Prandi J., Arbues A., Astarie-Dequeker C., Guilhot C., Werts C., Winter N, & Demangel C. (2018). Mycobacterial Phenolic Glycolipids Selectively Disable TRIF-Dependent TLR4 Signaling in Macrophages. Frontiers in Immunology, 9, 2.
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