Differential lymphoid-myeloid determination of leukocytes in whole blood

A lytic diluent comprising a mixture of ingredients (A) and (B) where

• (A) is an aqueous saline solution of at least one quaternary ammonium salt having surface active properties, and
• (B) is at least one additive selected from the group consisting of
• (1) a short chain alkanol substituted by phenyl or phenoxy;
• and
• (2) a polyhydroxy compound;
• said lytic diluent being buffered to an acid pH within the range of 3.5 to 5.0, and each of said ingredients (A) and (B) being present in an amount sufficient to bring about rapid, but incomplete, lysing of the red cells in whole blood for making a differential determination of lymphoid and myeloid populations of leukocytes, and also for measuring hemoglobin for chromagen formation, wherein said chromagen formation is completed by the addition of an alkaline reagent comprising an aqueous solution of a quaternary ammonium salt and an alkali metal cyanide, the mixture being buffered with a basic buffer in sufficient concentration to allow rapid neutralization of said lytic diluent which is buffered to an acid pH.

A method for the differential determination of lymphoid and myeloid populations of leukocytes and also for measuring hemoglobin by chromagen formation which comprises employing a diluent which is a mixture of ingredients (A) and (B) where

• (A) is an aqueous saline solution of at least one quaternary ammonium salt having surface active properties, and
• (B) is at least one additive selected from the group consisting of
• (1) a short chain alkanol substituted by phenyl or phenoxy;
• and
• (2) a polyhydroxy compound;
• said lytic diluent being buffered to an acid pH within the range of 3.5 to 5.0;
• and each of said ingredients (A) and (B) being present in an amount sufficient to bring about rapid and complete lysing of red cells in whole blood for making a differential determination of lymphoid and myeloid populations of leukocytes, and also for measuring hemoglobin by chromagen formation.

A method for the differential determination of lymphoid and myeloid populations of leukocytes and also for measuring hemoglobin by chromagen formation which comprises employing a diluent which is a mixture of ingredients (A) and (B) where

• (A) is an aqueous saline solution of at least one quaternary ammonium salt having surface active properties, and
• (B) is at least one additive selected from the group consisting of
• (1) a short chain alkanol substituted by phenyl or phenoxy;
• and
• (2) a polyhydroxy compound;
• said lytic diluent being buffered to an acid pH within the range of 3.5 to 5.0, and each of said ingredients (A) and (B) being present in an amount sufficient to bring about rapid, but incomplete, lysing of red cells in whole blood for making a differential determination of lymphoid and myeloid populations of leukocytes, and also for measuring hemoglobin by chromagen formation, wherein said chromagen formation is completed by the addition of an alkaline reagent comprising an aqueous solution of a quaternary ammonium salt and an alkali metal cyanide, the mixture being buffered with a basic buffer in sufficient concentration to allow rapid neutralization of said lytic diluent which is buffered to an acid pH.