Metachromatic dye sorption means for differential determination of leukocytes

The method of diagnostic aid in patient examination and disease treatment which comprises subjecting leukocytes of a patient donor blood specimen in a fixative-free state to supravital staining in an aqueous environment with the dye basic orange #21;

• microscopically comparing the donor leukocytes stained or non-stained thereby with a known non-diseased blood specimen similarly prepared being the base standard for comparison, noting differences from said standard in the tinctorial properties of the granules in the cytoplasm and nucleus present in said donor sample being diagnosed, thereby determining the presence or absence of disease affecting one or more of the leukocytes disclosing material modification from said standard in said donor specimen fraction.

A method of normal and pathologic human blood analysis whereby lymphocytes or leukemic lymphoblasts present in human blood specimen are made selectively spectrally determinant which comprises treating at least one supravital human blood specimen fraction in a fixative-free state by contact in an aqueous environment with at least one basic quaternary cationic dyestuff having the essential chromophoric combination of structure and groups present found in basic orange #21 thereby metachromatically differentiating the individual species of said cells from one another enabling classification and enumeration of identified species present in said analysis.

In a dye staining method for differential analysis of human blood specimen whereby mature leukocytes, immature leukocytes and other blood fragment components present in said specimen are made selectively determinant, one from the other, enabling differentiation and enumeration of all of said blood component species consisting essentially of neutrophils, eosinophils, basophils, lymphocytes, lymphoblasts and monocytes as well as myeloblasts, promyelocytes, myelocytes, metamyelocytes, platelets, bands, T-cells and B-cells;

• the method which comprises metachromatically staining a human blood containing specimen within a temperature range of 21.degree.-40.degree. C. in an aqueous, supravital, fixative-free state with substantially chemically pure, organic basic quaternary cationic dyestuff comprising basic orange #21;
• thereafter and thereby enabling identification, differentiation and enumeration of any one or all of the aforesaid blood component species present in said blood specimen under white light absorbance means.