Flow cytometry is a modern cell analysis technology that uses a flow cytometer to irradiate single cells or particles stained with fluorescent pigments under high-speed flow with high-energy lasers, measure the intensity of scattered light and emitted fluorescence, and thus qualitatively or quantitatively detect the physical, physiological, biochemical, immune, genetic, molecular biological characteristics and functional status of cells.
Experimental steps:
1. Use T25 culture flasks to culture each cell line. After the generation of cells grows to 80-90% fusion, discard the old culture medium
2. After washing the cells twice with 2ml of PBS, add trypsin to digest the cells. Observe the cells under the microscope until they become round. Add 4ml of complete culture medium to stop digestion
3. Gently blow off the cells with a pipette, transfer the cell mixture into a 15ml centrifuge tube, and centrifuge at 1000r/m for 10min. After the centrifugation, discard the culture supernatant, suspend the cell pellet with 5mlPBS, and centrifuge at 1000r/m for 10min
4. After discarding the supernatant, suspend the cell pellet with 500ul PBS. Then, antibodies were added and incubated at 4°C for 30 minutes for flow cytometry analysis
Results:
