Flow cytometry is a modern cell analysis technology that uses a flow cytometry sorter to irradiate single cells or microparticles stained with fluorescent pigments in a high-speed flow state with high-energy lasers, and measure the intensity of the scattered light and emitted fluorescence, so as to qualitatively or quantitatively detect the physical, physiological, biochemical, immune, genetic, molecular biological characteristics and functional status of cells. It can separate the subpopulations of luminescent particles according to the fluorescence intensity and wavelength of the emitted light and can realize monoclonal sorting, identify, classify, quantify and separate cells in complex samples, and the sorted cells can be directly used for culture, transplantation, nucleic acid extraction, single-cell PCR amplification or in situ hybridization, etc.
Experimental steps:
1. Use T25 culture flasks to culture each cell line. When the generation of cells reaches 80-90% fusion, discard the old culture medium
2. Wash the cells twice with 2ml PBS, add trypsin to digest the cells, and observe the cells become round under the microscope. Add 4ml complete culture medium to stop digestion
3. Use a pipette to blow off the cells gently, transfer the cell mixture into a 15ml centrifuge tube, centrifuge at 1000r/m for 10min, discard the culture medium supernatant, suspend the cell pellet with 5ml PBS, and centrifuge at 1000r/m for 10min
4. After discarding the supernatant, suspend the cell pellet with 500ul PBS. Then add antibodies respectively, incubate at 4℃ for 30min, and perform flow cytometry analysis using a flow cytometer
Result case:
