Coulter epics xl

Manufactured by Beckman Coulter

Sourced in United States, Spain, Germany, France

The Coulter Epics XL is a flow cytometry system designed for cell analysis and sorting. It utilizes the Coulter principle to measure and analyze the physical and chemical characteristics of particles, including cells, in a fluid suspension.

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Lab products found in correlation

Propidium iodide, by Merck Group (4 mentions) Modifit software, by BD (3 mentions) Facscalibur, by BD (3 mentions) Annexin 5 fitc 7 aad kit, by Beckman Coulter (2 mentions) P4170, by Merck Group (2 mentions) Annexin 5 fitc propidium iodide staining assay, by Thermo Fisher Scientific (2 mentions) Annexin 5 fitc, by Immunological Sciences (2 mentions) System 2 software, by Beckman Coulter (2 mentions) Cytomics fc500, by Beckman Coulter (2 mentions) Rnase free dnase, by Roche (2 mentions) Dihydroethidium, by Merck Group (2 mentions) Tcs sp2 confocal microscope, by Leica (2 mentions) Epics xl, by Beckman Coulter (2 mentions) Cxp software, by Beckman Coulter (2 mentions) Annexin 5 fitc, by BD (2 mentions) Ficoll paque plus, by GE Healthcare (2 mentions) Cobas ampliprep cobas taqman hiv 1 test, by Roche (2 mentions) Facscanto 2, by BD (1 mentions) Cell quest, by Beckman Coulter (1 mentions) Costar 6 well culture plates, by Corning (1 mentions)

Spelling variants of this product

Epics XL (474 citations) Coulter Epics XL flow cytometer (85 citations) Coulter Epics XL-MCL (25 citations) Coulter Epics XL/Xl-MCL (7 citations) Coulter Epics XL Flow Cytometry System (4 citations) Coulter Epics XL.MCL Cytometer (3 citations) Coulter Epics XL-MCLTM (3 citations) Coulter Epics XL FACS system (2 citations) Coulter Epics XL Flow Cytometric System (2 citations) Coulter Epics XL 4C (2 citations)

99 protocols using coulter epics xl

Flow Cytometric Analysis of RCC Cells

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Anti-human CD40-FITC and β1-integrin-FITC conjugated antibody with their isotype control (BD Biosciences, San Jose, CA, USA) were used for immunofluorescent staining of RCC cell lines. Cells were washed and re-suspended in FACS buffer (phosphate- buffered saline pH 7.2, 0.2% bovine serum albumin, and 0.02% sodium azide) containing 3% human serum and incubated with fluorochrome-conjugated antibodies for 15 min at 4 °C, then washed with the same buffer before flow cytometric analysis.
Cell fluorescence was acquired using a Coulter EPICS XL flow cytometer and analyzed using WinMDI version 2.9 software.
For apoptotic analysis 1 × 10⁶ cells were double stained with 20 μL of 7-AAD Viability Dye and 10 μL of Annexin V-FITC for 15 min on ice in the dark (Annexin V-FITC/7-AAD Kit, Beckman Coulter, Fullerton, CA, USA) and then analyzed on a Coulter EPICS XL equipped with the SYSTEM II software. Annexin V-FITC and 7-AAD emissions were detected in the FL-1 (525 nm) and FL-4 (675 nm) channels, respectively, and their bi-parametric histogram has shown the presence of four distinct populations: the viable cells, the apoptotic cells, the secondary necrotic cells and necrotic cells.

Pontrelli P., Gigante M., Spadaccino F., Netti G.S., Saldarelli M., Balducci L., Gigante M., Battaglia M., Storkus W.J., Castellano G., Stallone G., Gesualdo L, & Ranieri E. (2021). CD40 Cross-Linking Induces Migration of Renal Tumor Cell through Nuclear Factor of Activated T Cells (NFAT) Activation. International Journal of Molecular Sciences, 22(16), 8871.

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Apoptosis Quantification in Liver Cells

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The liver samples from different groups were thawed and minced with a scalpel in a cold PBS solution, incubated for 15 min at 4°C. The samples were filtered through a nylon mesh. After washing in the PBS solution and centrifugation, the cells chosen for analysis were collected and incubated with a solution containing propidium iodide (PI) (10 μg/ml, Sigma) and RNase (1 mg/ml, Sigma). The tubes were placed at 4°C in the dark for at least 30 min before analysis by flow cytometry. The PI fluorescence of individual nuclei was measured using Coulter Epics XL. At least 5 × 10³ cells of each sample were measured. Apoptotic and stained cells were represented by a subdiploid peak of cells that can be easily discriminated from the peak of cells with the diploid DNA content in the red fluorescence channel, analyzed by six-color flow cytometry on a FACS Canto II (BD Biosciences) with Flow Jo software (Tree Star, Inc., Ashland, OR). The percentage of apoptosis was indicated by the percentage of cells with subdiploid DNA content [27 ].

Al-Ghamdi T.H., Atta I.S, & El-Refaei M. (2020). Role of interleukin 6 in liver cell regeneration after hemi-hepatectomy, correlation with liver enzymes and flow cytometric study. Clinical and Experimental Hepatology, 6(1), 42-48.

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Apoptosis Induction Quantification

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Apoptosis induction was assessed by quantitation of histone-associated DNA fragment release into the cytosol using an ELISA kit from Roche Applied Sciences (Indianapolis, IN) or by flow cytometry using Annexin V/propidium iodide (PI) Apoptosis Detection kit. Quantitation of histone-associated DNA fragment release into the cytosol was performed according to the manufacturer’s instructions. For quantitation of apoptosis by flow cytometry, cells were treated with DMSO or SFN for 24 hours. Cells were harvested and washed with PBS. Cells were suspended in binding buffer and stained with Annexin V and PI for 15 minutes at room temperature in the dark. Samples were then diluted with binding buffer. Stained cells were analyzed using Coulter Epics XL or Accuri C6 Flow Cytometer.

Hahm E.R., Singh K.B., Kim S.H., Powolny A.A, & Singh S.V. (2020). The Role of Lysosome-Associated Membrane Protein 2 in Prostate Cancer Chemopreventive Mechanisms of Sulforaphane. Cancer prevention research (Philadelphia, Pa.), 13(8), 661-672.

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Phenotypic Characterization of Spleen Cells

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Spleen cells (1 × 10⁶ cells/mL) were blocked with anti-CD16/32 (FcγII/III receptor) mAb for 30 min at 4°C and then washed with PBS solution containing 1% FBS and 0.1% NaN₃. The cells were stained with PE-conjugated anti-CD8 mAb and FITC-conjugated anti-CD4 mAb or FITC-conjugated anti-CD19 mAb for 30 min at 4°C. Stained cells were then washed and detected by a flow cytometer (COULTER Epics XL, USA).

Kim J.J., Choi J., Lee M.K., Kang K.Y., Paik M.J., Jo S.K., Jung U., Park H.R, & Yee S.T. (2014). Immunomodulatory and Antidiabetic Effects of a New Herbal Preparation (HemoHIM) on Streptozotocin-Induced Diabetic Mice. Evidence-based Complementary and Alternative Medicine : eCAM, 2014, 461685.

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Cell Cycle Analysis by Flow Cytometry

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The cells were plated at a density of 5 × 10⁵ cells/dish. The day after, the cells, excluding the control samples, were treated either with 100 µM of APE (24 and 48 h of treatment) or with nocodazole (0.2 µg/mL) used as a positive control (24 h treatment). At the end of the treatments, cells were collected by trypsinization and fixed in PBS/ethanol (1:1; v/v). Samples were then incubated with monoclonal antibodies against phospho-histone H3 (p-HH3) (Ser10) (Merck Millipore, Darmstadt, Germany) for 60 min at room temperature, washed twice with 0.5% Tween-20 in PBS and incubated for 30 min with anti-mouse Alexa fluor 488-conjugated antibody (Invitrogen, Monza, Italy). Samples were washed with PBS and finally stained with 20 µg/mL propidium iodide for 15 min at room temperature. Flow cytometry analysis was performed using a flow cytometer Coulter Epics XL with 488 nm wavelength excitation. For each sample, at least 10⁴ events were measured.

Di Bari M., Tombolillo V., Alessandrini F., Guerriero C., Fiore M., Asteriti I.A., Castigli E., Sciaccaluga M., Guarguaglini G., Degrassi F, & Tata A.M. (2021). M2 Muscarinic Receptor Activation Impairs Mitotic Progression and Bipolar Mitotic Spindle Formation in Human Glioblastoma Cell Lines. Cells, 10(7), 1727.

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Cell Cycle Analysis by Flow Cytometry

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Nuclear DNA content was measured by propidium iodide (PI; Sigma-Aldrich) staining and fluorescence-activated cell sorting analysis according to the manufacturer's protocols. In brief, the cells were trypsinized and collected. Subsequent to three washes with phosphate-buffered saline (PBS), the cells were resuspended in 70% methanol and fixed in 4°C for 30 min. The cells were then washed and resuspended in PBS containing 20 µg/ml RNase A and 50 µg/ml PI, and incubated on ice for 30 min. Cell cycle analysis was performed in a Coulter Epics XL flow cytometer using the CellQuest program (Beckman Coulter, Inc., Brea, CA, USA) with manually set regions for the G₀/G₁, S and G₂/M phases. Data from 10,000 cells were collected for each data file.

XIONG Y., ZHANG L, & WANG T. (2015). Phosphorylation of BMK1 induces prostatic carcinoma cell proliferation by promoting entry into the S phase of the cell cycle. Oncology Letters, 11(1), 99-104.

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Cell Cycle and Apoptosis Analysis

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Cells were digested by 0.25% trypsin, seeded in six-well plates (5 × 10⁵ cells/well), and placed in nedaplatin solution at various concentrations (NDP1 and NDP2, see Section 2.3 above). For cell cycle analysis, cell culture was terminated after 24 h (Control, NDP1, and NDP2). For apoptosis analysis, cells were treated as described above (Control, RT, NDP1, NDP1 + RT, NDP2, and NDP2 + RT, see Section 2.3 above). Cells were collected by centrifugation at 1000 r/min, fixed with 70% ice-cold ethanol, adjusted to 1 × 10⁹/L, and stained with AnnexinV-fluorescein isothiocyanate (FITC)/propidium iodide (PI) for detecting apoptosis and PI for detecting cell cycle distribution (30 min, opaque background). Apoptosis and cell cycle distribution were detected by flow cytometry (Coulter EPICS XL, Coulter corp., USA) and data were analyzed by Multicycle software. The above procedures were also repeated three times.

Yin L., Wu J., Wu J., Ye J., Jiang X., Chen M., Wang D., Wang X., Zong D., Gu J., Zhang J., Wu J., Xu L., He X, & Guo W. (2014). Radiosensitization Effect of Nedaplatin on Nasopharyngeal Carcinoma Cells in Different Status of Epstein-Barr Virus Infection. BioMed Research International, 2014, 713674.

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Cell Cycle and Apoptosis Analysis

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Cell cycle phase distribution was analyzed after incubation with propidium iodide (PI, Sigma P4170). All parameters (FS, SS and FL-3) were acquired in a linear amplification scale. Cell aggregates were gated out on the bi-parametric graph FL-3lin/Ratio. Cell death was analyzed after incubation with Annexin V-FITC (Immunofluorescence Science, IK-11120), either alone or in combination with PI. In the latter case, both FL-1 (FITC) and FL-3 (PI) were acquired in a log amplification scale. Cell samples were analyzed in a Coulter Epics XL cytofluorometer (Beckman Coulter) equipped with EXPO 32 ADC software. At least 10,000 cells per sample were acquired. Data were processed using EXPO 32 ADC or Win MDI software.

Cesare E.D., Verrico A., Miele A., Giubettini M., Rovella P., Coluccia A., Famiglini V., Regina G.L., Cundari E., Silvestri R, & Lavia P. (2017). Mitotic cell death induction by targeting the mitotic spindle with tubulin-inhibitory indole derivative molecules. Oncotarget, 8(12), 19738-19759.

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Cell Cycle Analysis by Flow Cytometry

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Cells were plated onto Costar 6-well culture plates (Corning Incorporated, Corning, NY, USA) at 1.0×10⁶ cells/well and treated with 150 ng/ml rHuPCSK6. After 24 h, the cells were harvested by trypsinization, washed twice with phosphate-buffered saline (PBS), and fixed overnight with 70% ethanol at 4°C. The fixed cells were rinsed and resuspended in PBS, and stained for 30 min at 37°C with 1 ml 0.05 mg/ml propidium iodide solution containing 10 µg/ml RNase (Beijing Dingguo Biotechnology, Beijing, China). Cells were detected with a Coulter Epics XL flow cytometer, and DNA content was analyzed using Beckman Coulter EXPO32 ADC software (version 1.2B; both Beckman Coulter, Inc., Brea, CA, USA). The experiments were repeated 3 times in triplicate.

Wang P., Wang F., Wang L, & Pan J. (2018). Proprotein convertase subtilisin/kexin type 6 activates the extracellular signal-regulated kinase 1/2 and Wnt family member 3A pathways and promotes in vitro proliferation, migration and invasion of breast cancer MDA-MB-231 cells. Oncology Letters, 16(1), 145-150.

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Peripheral Blood Telomere Length Analysis

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Telomere length was determined in peripheral blood mononuclear cells (PBMC) by flow-FISH methodology by using the Telomere PNA kit/FITC from Dako (Glostrup, Denmark) according to the manufacturer's instructions. Samples were then analyzed by flow cytometry (Coulter Epics XL, Coulter, USA) and relative telomere length was determined by comparing isolated test cells with a control cell line (Human T-cell leukaemia 1301 cell line).

Pesi B., Ferrero A., Grazi G.L., Cescon M., Russolillo N., Leo F., Boni L., Pinna A.D., Capussotti L, & Batignani G. (2015). Liver resection with thrombectomy as a treatment of hepatocellular carcinoma with major vascular invasion: results from a retrospective multicentric study. American journal of surgery, 210(1).

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