Individual leukocyte determination by means of differential metachromatic dye sorption
First Claim
1. A dye sorbed and metachromatically dye stained, supra-vital, human blood cell specimen wherein minimally the monocytes species of leukocytes present in said dye sorbed specimen are differentiated, identified and enumerated by said dye sorption of color from among all other leukocytes present in said dye-sorbed specimen;
- said other leukocyte species comprising neutrophils, eosinophils, basophils and lymphocytes, said dye selected from the group of metachromatic dyes consisting of Griefswalder'"'"'s Blue, Blue Borrel, Rhodanile Blue, Toluylene Blue, Night Blue, Prune Pure, Hofmann'"'"'s Violet, Basic Orange 21, Basic Red 13, Basic Violet 16 and Carbocyanine K-5 and their admixtures;
said monocytes therein identified by the dye sorbed metachromatic differentiating color of their component nucleus and cytoplasm.
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Abstract
Lengthy cytochemical procedures for differentiation, identification, enumeration and diagnostic study of leukocytes in human blood have been advanced and shortened by a rapid method of optical differentiation of the five individual white blood cell species by selective use of basic quaternary metachromatic dye staining of supravital blood at controlled temperature. Presently a manual method, the improvement provides essential groundwork for improved accuracy in automated differential leukocyte counting.
36 Citations
16 Claims
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1. A dye sorbed and metachromatically dye stained, supra-vital, human blood cell specimen wherein minimally the monocytes species of leukocytes present in said dye sorbed specimen are differentiated, identified and enumerated by said dye sorption of color from among all other leukocytes present in said dye-sorbed specimen;
- said other leukocyte species comprising neutrophils, eosinophils, basophils and lymphocytes, said dye selected from the group of metachromatic dyes consisting of Griefswalder'"'"'s Blue, Blue Borrel, Rhodanile Blue, Toluylene Blue, Night Blue, Prune Pure, Hofmann'"'"'s Violet, Basic Orange 21, Basic Red 13, Basic Violet 16 and Carbocyanine K-5 and their admixtures;
said monocytes therein identified by the dye sorbed metachromatic differentiating color of their component nucleus and cytoplasm.
- said other leukocyte species comprising neutrophils, eosinophils, basophils and lymphocytes, said dye selected from the group of metachromatic dyes consisting of Griefswalder'"'"'s Blue, Blue Borrel, Rhodanile Blue, Toluylene Blue, Night Blue, Prune Pure, Hofmann'"'"'s Violet, Basic Orange 21, Basic Red 13, Basic Violet 16 and Carbocyanine K-5 and their admixtures;
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2. A composition of matter which comprises:
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(a) A leukocyte cell containing human blood specimen in a supra-vital fixative-free, aqueous environment reacted with (b) At least one pure, organic, basic quaternary cationic dyestuff;
said dyestuff differentiated from all other dyes in said class by the fact that in said foregoing composed environment all monocytes are stained metachromatically and differentially from all other blood cell leukocytes present in said dye stained composition. - View Dependent Claims (3, 4, 5)
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6. In a dye staining method for differential analysis of at least one of the leukocytes species consisting of neutrophils, eosinophils, basophils, lymphocytes and monocytes present in a human blood specimen;
- the improvement which comprises staining a supra-vital, aqueous human blood cell containing specimen within a temperature range of 21 to 40 C. with at least one substantially chemically pure, organic basic quaternary cationic dye;
all of the said dyes useful within said class characterized by their unique and unusual quality of differentially and metachromatically staining, minimally, the monocytes in said leukocyte group;
thereafter and thereby enabling the minimal identification, differentiation and enumeration of the said monocytes among said other leukocyte species in said blood specimen under white light absorbance means. - View Dependent Claims (7, 8, 9, 10, 11, 12, 13, 14, 15)
- the improvement which comprises staining a supra-vital, aqueous human blood cell containing specimen within a temperature range of 21 to 40 C. with at least one substantially chemically pure, organic basic quaternary cationic dye;
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16. A qualitative and quantative method of blood analysis for differential determination of blood leukocytes present in a supra-vital human blood specimen the steps which comprise:
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(a) Composing said blood specimen in a fixative free aqueous environment with (b) At least one organic basic quaternary cationic dyestuff;
said class of dyes identified by and restricted to those dyes in said class which stain monocytes metachromatically and differentially from all other leukocytes, said blood leukocytes consisting of neutrophils, eosinophils, basophils, lymphocytes and monocytes;
thereby inherently forming a differential dye-stained leukocyte cell-containing composition;
said composition uniquely useful thereafter in providing means to differentiate, identify and enumerate the so dyed individual leukocyte species present therein from all other leukocyte cells present in said metachromatically dyed cell differentiated blood cell containing composition by both automatic and manual cell differentiating means.
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Specification