Hello world,
I am currently doing an experiment with glioblastoma cells and cell killing using a small molecule drug. Here's how my supervisor told me to do it.
1. Serial dilute one 100 pi plate (~1*10^7 cells) to 7 dilutions (decreasing by half; 1*10^7, 1/2, 1/4, 1/8, 1/16, 1/32, 1/64) and later draw a standard curve on excel.
2.For the experimental group, seed 1/8 (from above) to 96 well plates.
3. Treat serially diluted drug for 2 days. Later draw a cell killing curve on excel
4. Treat alamarBlue reagent 1 day later, incubate 4 hours, read at 570nm/600nm
5. Find the slope equation of the standard curve
6. Punch in the absorbance values of the drug-treated cells into the equation to "normalize" the values
7. Get the percentage (100*x/control)
Here's my question, what does "normalize" in number 6 mean in this context? My supervisor told me that the values from the raw data do not reflect the actual % reduction of cells (or viability) and I am having trouble understanding...
Help would be appreciated!
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