Cell cycle analysis was determined using propidium iodide (PI) staining. Cells (1 × 107) were washed with PBS and fixed via 70% ethanol at room temperature for 30 min. After washing three times with PBS, cells were stained with PI contailing RNase A (Thermo, # F10797). Red signal was measured with a FACScan (Millipore). FSC data were analyzed using ModFIT LT v 3.1 software. For the apoptosis assay, MKN28 and AGS were staining by Annexin V and propidium iodide (PI) dual staining using an Annexin V-FITC Apoptosis Detection Kit (Vazyme, #A211-02). Fluorescence was measured with a FACScan (Millipore). FSC data were analyzed using Flowjo7.6 software.
Facscan
Manufactured by Merck Group
Sourced in United States, Germany
The FACScan is a flow cytometry instrument manufactured by Merck Group. It is used for the analysis and sorting of cells and particles in a fluid sample. The FACScan measures the physical and chemical characteristics of single cells or particles as they pass through a laser beam.
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Lab products found in correlation
8 protocols using facscan
1
Quantitative Cell Cycle and Apoptosis Analysis
2
Cell Apoptosis Assay via Annexin V-APC
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HO‐8910 and SK‐OV‐3 cells were seeded in six‐well plates and incubated at 37°C for 7 days. After incubation, the cells were harvested, centrifuged at 1300 rpm and washed using 4°C D‐Hanks (pH = 7.2 ~ 7.4). The cells were then resuspended in 200 μL of 1 × binding buffer and 10 μL of Annexin V‐APC (eBioscience) was added for 15 min in the absence of light. FACScan (Millipore) was applied to assess the apoptotic rate.
3
Annexin V-PI Apoptosis Assay
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Analysis of apoptosis on the indicated cell lines was performed by Annexin V/propidium iodide double staining followed by Flow cytometry (Guava Technologies, Millipore, USA). The cells 2 days after transfection were collected and subjected to analysis. A minimum of 5000 cells were then analyzed by FACScan with guavaSoft 3.1.1 software (Guava Technologies, Millipore, USA) for acquisition and analysis in three independent biological replicates.
4
Lentiviral Apoptosis Assay in Cell Lines
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Lentiviral Eca-109 and TE-1 cells transfected for 5 days were inoculated in culture dishes, digested with trypsin, suspended, and stained with Annexin V-APC (10 µL) in the dark for 15 min. Cell apoptosis rates were assessed using FACScan (Millipore, Darmstadt, Germany, USA). Meanwhile, the green fluorescence of GFP labeled on lentivirus was detected, as shown in the Y-axis. The cells not transfected with lentivirus were excluded. Therefore, the rate of apoptosis was accurately determined.
5
Apoptotic Rate Quantification in U87 and U251 Cells
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U87 and U251 cells (at least 5 × 105) were suspended and seeded in 6 cm dishes and cultured until cells covered approximately 70% of the plate. The cells were digested with trypsin and resuspended, then stained with the addition of 10 μL annexin V‐APC for 15 minutes in the dark. The percentages of cell phases were measured using FACScan (Millipore) to assess the apoptotic rate, and results were analyzed.
6
Cell Cycle Analysis via PI Staining
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Propidium iodide (PI) staining was utilized to analyze cell cycle. In short, for the cell cycle analysis, cells (1 × 107) were washed using PBS and fixed using 70% ethanol for 30 min at room temperature. We then stained the cells with PI containing RNase A (Thermo, # F10797) after washing them three times with PBS. A FACScan (Millipore) was used to measure the red signal and ModFIT LT v3.1 software was used to analyze the FSC data.
7
Apoptosis Assessment in Cancer Cells
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Infected HO-8910 and OVCAR-3 cells were harvested and centrifuged at 1300 rpm for 5 min, and the cell precipitation was washed with ice-cooled D-Hanks (pH=7.2-7.4). Then cells precipitation was suspended with 200 μL 1 × binding buffer and then cells were stained by 10 μL Annexin V-APC for 15 min in the dark. FACScan (Millipore) was used to assess the apoptosis rate.
8
Apoptosis and Cell Cycle Analysis
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Infected HCT116 and RKO cells were harvested and centrifuged (1300 rpm) for 5 min, and then washed with 4 °C pre-cooled D-Hanks. Then cells suspension with 5 × 105 cells using 1×binding buffer was stained by 10 µL Annexin V-APC for 15 min in the dark. FACScan (Millipore) was used to assess the apoptosis rate.
shCHSY1 and shMCM8 + shCHSY1 lentivirus infected RKO cells were cultured in a 6 cm dish for 5 days, and then cells were harvested and washed with PBS. After centrifuged (1200 rpm) for 5 min, cells were washed with 4 °C pre-cooled PBS and fixed by 70% ethanol for 1 h. After centrifuged at 1500 rpm, 1 mL PI staining solution was added for cell cycle distribution detecting using FACSCalibur (BD Biosciences) at 200 ~ 300 Cell/s.
shCHSY1 and shMCM8 + shCHSY1 lentivirus infected RKO cells were cultured in a 6 cm dish for 5 days, and then cells were harvested and washed with PBS. After centrifuged (1200 rpm) for 5 min, cells were washed with 4 °C pre-cooled PBS and fixed by 70% ethanol for 1 h. After centrifuged at 1500 rpm, 1 mL PI staining solution was added for cell cycle distribution detecting using FACSCalibur (BD Biosciences) at 200 ~ 300 Cell/s.
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