Formulation for total and differential counting of leukocytes in liquid medium and method of making and using same

A method of preparation and formulation of a composition for total and differential counting of leukocytes in liquid medium, comprising the following steps:

• A1) Step of production of a dye solution, with the dilution of cresyl acetate violet dye in anhydrous ethyl alcohol;
• then the resulting dilution is kept for 1 hour, protected from light;
• then filtered through filter paper No. 1 or No. 5;
• then deionized water is added to the dilution;
• A2) Step of producing a hemolyzing solution, with the dilution of sodium chloride (NaCl) and potassium chloride (KCl) in deionized water, followed by the addition of 1M sodium hydroxide solution (1 molar), glacial acetic acid and t-octylphenoxypolyethoxyethanol;
• then the solution is homogenized on a thermal magnetic plate heated to 50° C., until the complete dilution of all components;
• and
• A3) Step of preparation of the composition by mixing 1 part by volume of the dye solution with 9 parts by volume of the hemolyzing solution;
• buffering to a pH between 4.0 and 6.0 ration;
• and storing the composition protected from light.