محلول آماده تریپان بلو 10mL

This method will enable you to  determine the cell viability. Cell viability is calculated as the number of viable cells divided by the total number of cells within the grids on the hemacytometer. 

If cells take up trypan blue and staine blue, they consider as dead cells.

1.  Using a hemacytometer, the cell density of the cell line suspension should be determine. 
2. Add 100 ul of trypan blue stock solution (DBT005/DBT0010) to 100 ul of cells (1:1 ratio) (e.g into a 0.5 mL microtube).
3. (Optionl) the above reagent could be incubated in RT (2-3 min) or 37^o C (shorter period of time).
4. Transfer to a hemacytometer and examine immediately under a microscope at low magnification.
5. Count the number of blue cells and the number of total cells. 

% viable cells = [1.00 – (The number of blue staining cells ÷ the number of total cells)] × 100

To calculate the number of viable cells per mL of culture, use the formula below:

Number of viable cells × 10⁴ × 1.1 = cells/mL culture

Note: remember to correct for the dilution factor.