Cells were fixed with 70% ethanol. Briefly, cells were collected and washed in PBS–2%BSA twice and resuspended to a concentration of 3–6 mill/ml. 500 ml were taken and 5 ml cold 70% ethanol was added dropwise to the cells whilst gently vortexing to prevent cell clumping. Cells were fixed at 4 °C for 1 h and stored at − 20 °C until ready for FACS analysis. The cells were stained with FxCycle PI/RNase Staining Solution solution (Life Technologies F10797) according to the manufacturer’s protocol and the cells were analysed using the BD FACS Canto 11 system with the BD FACS Diva software version 8 (BD Biosciences, San Jose, CA, USA). Data analysis was performed using the Modfit4 software (Verity Software House, Topsham, ME, USA).
Facs canto 11 system
Manufactured by BD
Sourced in United States
The BD FACS Canto II system is a flow cytometry instrument designed for cell analysis and sorting. It features a three-laser configuration and can detect up to 10 parameters simultaneously. The system is capable of analyzing a wide range of sample types, including cells, particles, and microorganisms.
Automatically generated - may contain errors
2 protocols using facs canto 11 system
1
Ethanol-Based Fixation and PI Staining for Flow Cytometry
2
Cell Cycle Analysis via Flow Cytometry
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Cells were fixed with 70% ethanol. Briefly, cells were collected and washed in PBS–2%BSA twice and resuspended to a concentration of 3–6 mill/ml. 500 ml were taken and 5 ml cold 70% ethanol was added dropwise to the cells whilst gently vortexing to prevent cell clumping. Cells were fixed at 4 °C for 1 h and stored at −20 °C until ready for FACS analysis. The cells were stained with FxCycle PI/RNase Staining Solution solution (Life Technologies F10797) according to the manufacturer’s protocol and the cells were analysed using the BD FACS Canto 11 system with the BD FACS Diva software version 8 (BD Biosciences, San Jose, CA, USA). Data analysis was performed using the Modfit4 software (Verity Software House, Topsham, ME, USA).
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