METACHROMATIC DYE SORPTION AND FLUORESCENT LIGHT EMISSIVE MEANS FOR DETERMINATION OF DEVELOPMENTAL STAGES OF NEUTROPHILIC GRANULOCYTIC CELLS AND OTHER LEUKOCYTES

摘要

TITLE: METACHROMATIC DYE SORPTION ANDFLUORESCENT LIGHT EMMISIVE MEANSFOR DIFFERENTIAL DETERMINATION OFDEVELOPMENTAL STAGES OF NEUTROPHILICGRANULOCYTIC CELLS AND OTHER LEUKOCYTESABSTRACT OF THE INVENTIONIn the diagnosis and prognosis of various diseases aswell as medical researches in relation thereto studies leadingto differentiation, identification, enumeration and followingthe variations in human blood, determination of developmentalstages of neutrophilic granulocytic cells including promyelo-cytes, myelocytes, metamyelocytes, bands, neutrophils,eosinophils, basophils, monocytes, B-lymphocytes or B-cellsand T-lymphocytes or T-cells and other leukocytes play a vitalrole. Heretofore, accurate detailed lengthy and arduous pro-cedures were essential to such ends, if available. In aParent case, Canadian application number 371180, filed February 18, 1981it was found selective use of certain basic quaternary, meta-chromatic dye staining by supravital techniques made possibleinstantaneous optical differentiation of the five majorindividual leukocyte species. In Canadian applicationNo. 397322 filed March 1, 1982,it was disclosed that with one sub-set ofspecies of the above dye class one can by the same whitelight wave spectra techniques make the foregoing determinationswith regard to specific leukocytes including promyelocytes,myelocytes, metamyelocytes, band cells, T-cells and B-cellsas well as the neutrophils, eosinophils, basophils and mono-cytes of the Parent disclosure.Now it is known that by similar observations as above,but under fluorescent light emission from the dyed specimen,similar differentiation, identification and enumeration ofbasic orange #21 dyed supravital cells as listed can beaccomplished. Under both absorbance and fluorescence thesame physical structural elements are generally useful andare not appreciably altered. Patterns of granular distributionand quality are not appreciably altered. However, the colorsunder fluorescent light response are dissimilar and enoughso that no confusion as to identity with white light absor-bance results. As one uses both modes of examination of thesame sample analyses can be confirmed accurately.Using coherent laser light in stimulating the fluores-cent mode suggests application to flow cytometry methods ofsimilar analysis by automatic equipment. The bi-modal lightsources offer use of cytological comparisons which revealdetails of comparisons and differences not heretofor recorded.Identified basic reds and violets of the methine andpolymethine chromophoric class cause metachromatic stainingof monocyte nucleus in absorbtive light as well as undermetachromatic fluorescence thereby verifying monocyte identi-fication.