The cells were treated as described above, then washed with PBS and resuspended in the binding buffer containing Annexin V-FITC and PI for 15 min in the dark. Thereafter, the cells were detected by Navios flow cytometry (Beckman Coulter, Brea, CA, USA). The percentage of apoptotic cells was counted using Navios software (Beckman Coulter, Brea, CA, USA).
Navios flow cytometry
Manufactured by Beckman Coulter
Sourced in United States
The Navios flow cytometry system is a high-performance instrument designed for multi-color flow cytometry analysis. It enables the simultaneous detection and measurement of multiple cellular parameters, such as size, granularity, and the expression of specific proteins or markers on the surface or within cells. The Navios system is capable of analyzing a wide range of sample types, including cells, microparticles, and other biological particles.
Automatically generated - may contain errors
Lab products found in correlation
Navios software, by Beckman Coulter (2 mentions)
7 aminoactinomycin d 7 aad, by Thermo Fisher Scientific (1 mentions)
Celltrace violet cell proliferation kit, by Thermo Fisher Scientific (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Kaluza software 2, by Beckman Coulter (1 mentions)
Cell cycle reagent, by Beckman Coulter (1 mentions)
Annexin 5 fitc apoptosis detection kit, by Beckman Coulter (1 mentions)
Tacs annexin 5 fitc apoptosis detection kit, by R&D Systems (1 mentions)
Dna prep reagent kit, by Beckman Coulter (1 mentions)
Wincycle32, by Beckman Coulter (1 mentions)
Tacs annexin 5 fitc, by Bio-Techne (1 mentions)
Kaluza software, by Beckman Coulter (1 mentions)
Annexin v 7aad apoptosis assay kit, by BD (1 mentions)
13 protocols using navios flow cytometry
1
Quantifying Apoptosis via Flow Cytometry
2
Mesenchymal Stem Cell Phenotyping Assay
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To assess the MSC phenotype, cytometric analysis was performed using Dura clone SC Mesenchymal Tube antibodies (Beckman Coulter, India, Bangalore, Karnataka). DURA Clone MSC tubes contain lyophilisate of a panel of 9 monoclonal antibodies (CD90-FITC, CD73-PE, CD34-ECD, CD146-PC5.5, CD105-PC7, CD45-APC-AF750, CD31-Pacific Blue, CD14-Krome Orange, CD19-Krome Orange) dedicated to the determination of characteristic MSC antigens.10 (link),26 (link),27 (link) Cell proliferation was assessed using the anti Ki67-PE eBioscience antibody (Invitrogen by Life Technologies, USA, Carlsbad, CA) and viability of the cultured cells using propidium iodide and annexin V. Staining with annexin-V, which can bind to phosphatidylserine on the surface of apoptotic cells, was used as a marker of apoptosis. Annexin V was detected by flow cytometry in FITC and PI channels. For this purpose, a commercially available annexin-V staining detection kit was used to measure apoptosis and cell viability (Invitrogen, USA, Eugene, Oregon). MSC immunophenotyping was performed on a Navios flow cytometry (BeckmanCoulter). The cytometric analysis procedure of Ki67 and annexin V is described in the work Fathi et al.28 (link)
3
T cell-mediated cytotoxicity assay
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CD30+ cell lines, K562 cell lines, and CD30- cell line, SupB15, were stained with 1μM of CellTrace™ Violet cell proliferation kit (Life Technologies Corporation, ThermoFisher Scientific, USA) and co-cultured individually with engineered- and mock T cells at effector-to-target ratios (E: T) of 1:1 and 40:1. After 24 hours of culture, the cell pellet was stained with 7-amino-actinomycin D (7AAD) (eBioscience, USA) to detect the proportion of dead target cells and analyzed by Navios Flow Cytometry (Beckman Coulter, USA) and FlowJo software (version 10, TreeStar, USA). The specific lysis was calculated as follows;
4
Monitoring Immune Cell Populations in ABOi LDLT
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The cluster of differentiation (CD) marker levels (%), including CD 4+, 8+, 19+, 20+, and 25+ were estimated 1 day before surgery and on POD 14 in patients undergoing ABOi LDLT. Blood samples were obtained in test tubes (BD Vacutainer, K2 EDTA; Becton, Dickinson and Company, Franklin Lakes, NJ, USA) and sent to the laboratory for analysis of the CD markers using Navios flow cytometry (Beckman Coulter, Indianapolis, IN, USA).
5
Apoptosis and Cell Cycle Analysis
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Flow cytometry was used to study cell apoptosis and cell cycle after treatment with different compounds. Cells (5 × 105/well) were plated in 6-well plates and incubated for 24 h in the presence or absence of MA1. An apoptosis assay was performed using Annexin V-FITC Apoptosis Detection Kit (PN IM3546, Beckman Coulter, Indianapolis, IN, USA), while cell cycle reagent (C03551, Beckman Coulter, Indianapolis, IN, USA) was used to assess the cell cycle distribution pattern for A2780S and A2780CP cells, according to the manufacturer’s protocol. Briefly, cells were detached using 0.05% Trypsin/EDTA (Gibco, New York, NY, USA) and, for apoptosis analysis, Annexin V and PI dyes were added and incubated in the dark on ice for 20 min. In the cell cycle, cells were harvested and immediately fixed in 70% (v/v) ethanol overnight at 4 °C, then stained with PI reagent. The data were acquired using Navios flow cytometry (Beckman coulter Life Sciences, Indianapolis, IN, USA) and analyzed using Kaluza software 2.1 (Beckman coulter Life Sciences, Indianapolis, IN, USA).
6
Apoptosis Assessment in Granulosa Cells
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To determine the effects of H2O2–KLF12 on the induction of apoptosis in GCs, the TACS Annexin V-FITC Apoptosis Detection Kit (R&D Systems, Inc) was used according to the manufacturer’s guidelines. Briefly, GCs were digested with trypsin–EDTA. The cells were collected, gently washed with PBS, and counted. Cells were collected by centrifugation at approximately 300g for 5 min at room temperature and resuspended in Annexin V Incubation Reagent at a concentration of 1 × 106 cells/100 μl. Cells were incubated in the dark for 15 min at room temperature. Finally, 400 μl of 1× binding buffer was added to stained cells and acquired by Navios flow cytometry (Beckman Coulter Life Science) within 1 h for maximal signal. The results were analyzed by Navios software (Beckman). Finally, 10 μl propidium iodide (PI) was added for cell flow cytometry analysis. Viable cells (Annexin V−/PI−) do not take any color, early apoptotic cells (Annexin V+/PI−) are green, late apoptotic cells (Annexin V+/PI+) are green and orange, and necrotic cells (Annexin V−/PI+) are orange.
7
Lymphocyte Immunophenotyping by Flow Cytometry
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Flow cytometry immunophenotyping was used to count CD19+ lymphocytes. At least 5,000 lymphocytes were analyzed by Navios flow cytometry (Beckman Coulter, Miami, FL). The analysis was stopped when a minimum of 20 CD19+ events were detected. The maximum number of lymphocytes analyzed was 200,000. Lymphocyte counting was planned every 6 months.
8
Cell Cycle Analysis by Flow Cytometry
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The cell cycle analysis was performed using a DNA Prep™ reagent Kit (Beckman Coulter, Inc.) by NAVIOS flow cytometry (Beckman Coulter, Inc.) according to the manufacturer’s protocol. The fractions of the cell population in the G1/G0 (G01), S, and G2/M phases were quantified via using the Wincycle32 software (Beckman Coulter, Inc.). The sub-G1 fraction was determined from the total event count.
9
Quantifying Circulating Plasma Cells in Multiple Myeloma
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Ethylenediaminetetraacetic acidperipheral blood (EDTA-PB) samples were collected from 145 MM patients when diagnosed, and 10-color flow cytometry was evaluated within 24 h after collection. Samples were labeled with antibodies CD138-APC/CD38-APC750/CD45-KO/CD19-ECD/CD56-PC7/CD27-PB/CD81-APC700/CD117-PC5 (Beckman Coulter, Beijing, China) and cytoplasmic Kappa and Lambda immunoglobulin light chains (Dako, Shanghai, China). Beckman Coulter Navios flow cytometry was used for analysis. The gating strategy was applied with the level of CD38 and CD138 to identify all plasma cells. The cPCs were identified based on Kappa and that chains restricted expression. At least 50,000 nucleated cells were analyzed for each tube. The percentage of CPC was expressed as CPC/total nucleated cells from the blood. CPC negativity was defined as the absence of CPC in the blood with a limit of detection of <1×10−4, while CPC positivity was defined as the level of CPC higher than this threshold.
10
Apoptosis Detection in Dendritic Cells
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To determine the effects of BSEO treatment on the induction of apoptosis or necrosis in DCs, apoptosis detection kit (TACS®Annexin V-FITC, Trevigen, Gaithersburg, USA) was used and performed according to the manufacturer’s guidelines. CPT was used as an apoptosis inducer at a concentration of 10 mM for 4 h and purchased from Sigma Aldrich®,St. Louis, USA. Briefly, cells were collected, washed once, and resuspended in 100 μL of Annexin-V reagent. Cells were left in the dark for 15 min at room temperature. Finally, 400 μL of 1X binding buffer was added to stained cells and acquired by Navios flow cytometry (Beckman coulter life science, USA) within 1 h and analyzed using Navios software [20 (link)]. Ten μL of PI was added to the cells prior to analysis by flow cytometry. Consequently, viable cells do not take any color (Annexin V−/PI-), early apoptotic cells are green (Annexin V+/PI-), late apoptotic cells are green and orange (Annexin V+/PI+), and necrotic cells are orange (Annexin V−/PI+).
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