Collect cultured cells and centrifuge at 300× gravity for 5 minutes to pellet the cells. Discard the supernatant.
▶ 00:16
2
Count cells and adjust concentration
Resuspend cells in fresh calcium medium and count them using a microscope. Adjust the cell concentration as needed.
▶ 00:41
3
Set up experimental plate and controls
Prepare the 96-well plate by adding 100 µL PBS to blank wells, 100 µL calcium medium to negative control wells, and 100 µL cell suspension to experimental wells according to the experimental design.
▶ 01:31
4
Treat cells and incubate
Apply experimental treatments to the cells according to the experimental protocol and incubate in a cell incubator.
▶ 01:59
5
Add CCK-8 reagent and incubate
Add 10 µL of CCK-8 buffer to each well and incubate the plate to allow the color reaction to develop.
▶ 02:49
6
Measure optical density values
Measure the optical density (OD) value of each well at 450 nanometers using a microplate reader.
▶ 03:29
7
Calculate and analyze data
Organize data by group and calculate average OD values. Subtract the blank group average from each group, then divide by the negative control value minus blank to determine the cell viability rate.