Pyroptosis rate in the specimens was determined using a FAM-FLICA Caspase-1 detection kit (ImmunoChemistry) (Zeng et al. 2019 ). Brie y, prepared cells were stained with FAM-FLICA (10 μl) and PI (5 μl) at 26℃ for 20 minutes. Then, uorescence intensities of the specimens were measured by a Coulter Epics XL ow cytometer. Pyroptosis rate was calculated using a formula: number of doublepositive cells / number of total cells × 100%.
Epics xl ow cytometer
Manufactured by Beckman Coulter
The Epics XL flow cytometer is a laboratory instrument designed to analyze and measure the physical and chemical characteristics of cells or particles suspended in a fluid stream. It utilizes laser technology to detect and differentiate between various cell types based on their size, granularity, and fluorescent properties.
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Lab products found in correlation
5 protocols using epics xl ow cytometer
1
Pyroptosis Rate Quantification
2
Pyroptosis Rate Quantification
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Pyroptosis rate in the specimens was determined using a FAM-FLICA Caspase-1 detection kit (ImmunoChemistry) (Zeng et al. 2019 ). Brie y, prepared cells were stained with FAM-FLICA (10 μl) and PI (5 μl) at 26℃ for 20 minutes. Then, uorescence intensities of the specimens were measured by a Coulter Epics XL ow cytometer. Pyroptosis rate was calculated using a formula: number of doublepositive cells / number of total cells × 100%.
3
Pyroptosis Quantification by Flow Cytometry
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Pyroptosis rate was determined by a FAM-FLICA in vitro caspase-1 detection kit (ImmunoChemistry, USA) [23] . First, the cells were mixed with trypsin. Second, the cells were washed several times using phosphate buffer saline. Third, the cells were stained with 10 µl FAM-FLICA and 5 µl PI for 20 minutes in a dark environment at room temperature. Fourth, uorescence intensity was measured using a Coulter Epics XL ow cytometer. Pyroptosis rate was calculated using a formula: number of double-positive cells / number of total cells × 100%.
4
Cell Cycle Analysis via Flow Cytometry
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Constructed transfectants were cultured in each well of six-well plates to 70-80% con uence using normal culture medium. The cell culture medium was replaced with 0.1% FBS DMEM with 2 mmol/L Lglutamine and 25 µg gentamicin, and the constructed transfectants were again cultured for 24 h. Cells were then suspended in 70% ethanol for 24 h at 4 °C, then incubated with RNase A at 37 °C for 30 min, and stained with propidium iodide (PI) at 4 °C for 30 min. DNA content was determined by ow cytometry using the Epics XL ow cytometer (Beckman Coulter Inc., San Diego, CA), as previously described(18).
5
Cell Cycle Analysis via Flow Cytometry
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Constructed transfectants were cultured in each well of six-well plates to 70-80% con uence using normal culture medium. The cell culture medium was replaced with 0.1% FBS DMEM with 2 mmol/L Lglutamine and 25 µg gentamicin, and the constructed transfectants were again cultured for 24 h. Cells were then suspended in 70% ethanol for 24 h at 4 °C, then incubated with RNase A at 37 °C for 30 min, and stained with propidium iodide (PI) at 4 °C for 30 min. DNA content was determined by ow cytometry using the Epics XL ow cytometer (Beckman Coulter Inc., San Diego, CA), as previously described(18).
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