After 16 h of VSV-GFP infection, the indicated cells were harvested and resuspended in 0.5 mL of phosphate-buffered saline (PBS). The samples were analyzed by flow cytometry (Beckman Coulter, Fullerton, CA, USA), and the flow cytometry data were analyzed using FlowJo software (TreeStar, Ashland, OR, USA).
Flow cytometry
Manufactured by Tree Star
Sourced in United States
Flow cytometry is an analytical technique used to measure and analyze the physical and chemical characteristics of particles, cells, or other biological entities. It is a powerful tool for a wide range of applications, including cell counting, cell sorting, biomarker detection, and more. The core function of flow cytometry is to rapidly analyze multiple parameters of single cells or particles as they flow in a fluid stream through a laser beam or other interrogation point.
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Lab products found in correlation
Apoptosis assay kit, by Thermo Fisher Scientific (1 mentions)
Annexin 5 apc, by BD (1 mentions)
Annexin 5, by BD (1 mentions)
7 aad, by BD (1 mentions)
Anti cd25 pe cy7, by Thermo Fisher Scientific (1 mentions)
Anti cd4 fitc, by Thermo Fisher Scientific (1 mentions)
Anti cd8 pe, by Thermo Fisher Scientific (1 mentions)
Apc anti cd69, by Thermo Fisher Scientific (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Anti cd3 percp, by Thermo Fisher Scientific (1 mentions)
Annexin 5 fitc, by BD (1 mentions)
Propidium iodide, by BD (1 mentions)
Annexin 5 fitc pi apoptosis kit, by BD (1 mentions)
Flow cytometry, by BD (1 mentions)
7 protocols using flow cytometry
1
Flow Cytometry Analysis of VSV-GFP Infection
2
Apoptosis Assay of ABGE-Treated Cells
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The apoptosis assay kit (Invitrogen, USA) was used for cell apoptosis analysis. In 24 well culture plates, cells were seeded (about 5*105 cells/well) and then incubated for 24 h. The cells were then given 100mg/ml ABGE or culture media (negative control). Apoptosis was then examined by Annexin V/PI staining, followed by flow cytometry analysis of stained cells. ABGE-treated cells were harvested after 48 hours, washed twice with cold PBS, and resuspended in 1X Annexin binding buffer. 6ul FITC-conjugated Annexin V and 1ul PI were added to a fresh 1.5 ml tube containing 100 ml of binding buffer and cells. In each tube, 200 ml of 1X Annexin binding buffer was added after 15 minutes of incubation. The stained cells were analyzed by flow cytometry (BD Biosciences, USA) with flowjo software (v10.6.2, Treestar, USA).
3
Apoptosis Evaluation via Annexin V-FITC/7-AAD
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Fluorochrome-labeled Annexin V (Annexin V-FITC) was used to specifically target and identify early apoptotic cells. Late apoptotic cells, which have lost membrane integrity, are recognized by 7 -aminoactinomycin D (7-AAD). In contrast, necrotic cells lose membrane integrity early and become 7-AAD positive. The prepared cells were plated in 96-well plates at 1 × 106 cells each, seeding 200 μL cell suspensions per well, and then treated or untreated with the concentrations as described in the text of CsA for 48 h with 5% CO2 at 37 °C. Cells were washed once with PBS and resuspended in 500 μL PBS. Then, 200 μL of cell suspensions were transferred into a tube and then incubated with 1.5 μL Annexin V and 1 μL 7-AAD (BD Biosciences, San Jose, CA, USA) for 30 min. After supernatants were discarded via centrifugation at 1500 rpm/min for 5 min, the cells were resuspended with 200 μL binding buffer and stained with 1 μL APC-Annexin V (BD Biosciences) for 15 min. The apoptosis analysis was performed using flow cytometry (TreeStar, Ashland, OR, USA).
4
Immunophenotyping of Lymphocytes in PCOS
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This experiment was performed following previously described procedures29 (link). Single-cell suspensions from the follicular fluid of infertile women with and without PCOS were adjusted to 0.3 × 106/ml, washed twice in phosphate-buffered saline (PBS) (Invitrogen Life Technologies, Grand Island, NY, USA) and blocked in PBS buffer that contained 1% bovine serum albumin (BSA) (Sigma-Aldrich, St. Louis, MO, USA) for 30 min. The cells were then stained for 30 min at 4 °C in the dark with conjugated antibodies specific for the following cell surface antigens: anti-CD3 PerCP, anti-CD4 FITC, anti-CD8 PE, anti-CD25 PE-CY7, anti-CD69 APC and anti-PD-1 Brilliant Violet 421 (eBioscience, San Diego, CA, USA). The phenotypic characteristics of the antibody-labeled lymphocytes were analyzed using flow cytometry (Beckman Coulter, Fullerton, CA, USA), and the results were analyzed using FlowJo version 6.0 software (TreeStar Inc., Ashland, OR, USA). Isotype-matched controls were included in each staining protocol.
5
Measuring Apoptosis by Flow Cytometry
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The serum-starved cells were prepared and cultured as described above. Following treatment with AG490 at the indicated concentrations for 24 h, cells were washed with cold PBS 3 times, and then incubated with Annexin V-FITC and PI at room temperature in the dark for 15 min. Samples were analyzed by flow cytometry (BD Biosciences) within 1 h. Data were analyzed using FlowJo7.6 software (Tree Star, Inc.).
6
Butorphanol-Induced Apoptosis Analysis
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5 x 105 cells were cultured in each well of a 6-well plate, and incubated in different concentrations of Butorphanol for 48 h. Then, cells were harvested and resuspended in binding buffer. The cells were stained with 5 μL Annexin V-FITC plus 5 μL propidium iodide (BD Biosciences, Franklin Lakes, NJ). The stained cells were added of 400 μL PBS. The percentage of apoptosis was analyzed by using flow cytometry (Tree Star, Ashland, OR).
7
Quantifying Apoptosis in OSCC Cells
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Flow cytometry (BD Biosciences, USA) was performed to assess the cell apoptosis ability of transfected OSCC cells using the Annexin V-FITC/PI Apoptosis kit (Cat#: 556,547; BD, USA) according to a previous study [31 (link)]. After 48 h transfection, 1 × 106 SCC-4 and HSC-3 cells were collected in 100 μL of binding buffer. Then, the cells were incubated with 10 μL Annexin V-FITC and 10 μL propidium iodide (PI) at 22–25°C in the dark. The cell apoptosis rate was detected by Flow cytometry and analyzed by FlowJo 7.6.1 (Treestar, USA).
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