G - Physics – 01 – N
Patent
G - Physics
01
N
150/11, 73/51.5
G01N 21/64 (2006.01) G01N 33/50 (2006.01) G01N 1/30 (2006.01)
Patent
CA 1205365
TITLE: METACHROMATIC DYE SORPTION AND FLUORESCENT LIGHT EMMISIVE MEANS FOR DIFFERENTIAL DETERMINATION OF DEVELOPMENTAL STAGES OF NEUTROPHILIC GRANULOCYTIC CELLS AND OTHER LEUKOCYTES ABSTRACT OF THE INVENTION In the diagnosis and prognosis of various diseases as well as medical researches in relation thereto studies leading to differentiation, identification, enumeration and following the variations in human blood, determination of developmental stages of neutrophilic granulocytic cells including promyelo- cytes, myelocytes, metamyelocytes, bands, neutrophils, eosinophils, basophils, monocytes, B-lymphocytes or B-cells and T-lymphocytes or T-cells and other leukocytes play a vital role. Heretofore, accurate detailed lengthy and arduous pro- cedures were essential to such ends, if available. In a Parent case, Canadian application number 371180, filed February 18, 1981 it was found selective use of certain basic quaternary, meta- chromatic dye staining by supravital techniques made possible instantaneous optical differentiation of the five major individual leukocyte species. In Canadian application No. 397322 filed March 1, 1982,it was disclosed that with one sub-set of species of the above dye class one can by the same white light wave spectra techniques make the foregoing determinations with regard to specific leukocytes including promyelocytes, myelocytes, metamyelocytes, band cells, T-cells and B-cells as 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 of basic orange #21 dyed supravital cells as listed can be accomplished. Under both absorbance and fluorescence the same physical structural elements are generally useful and are not appreciably altered. Patterns of granular distribution and quality are not appreciably altered. However, the colors under fluorescent light response are dissimilar and enough so that no confusion as to identity with white light absor- bance results. As one uses both modes of examination of the same sample analyses can be confirmed accurately. Using coherent laser light in stimulating the fluores- cent mode suggests application to flow cytometry methods of similar analysis by automatic equipment. The bi-modal light sources offer use of cytological comparisons which reveal details of comparisons and differences not heretofor recorded. Identified basic reds and violets of the methine and polymethine chromophoric class cause metachromatic staining of monocyte nucleus in absorbtive light as well as under metachromatic fluorescence thereby verifying monocyte identi- fication.
420950
Kass Lawrence
Ridout & Maybee Llp
LandOfFree
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