Reagent compositions and their use in the identification and...

C - Chemistry – Metallurgy – 09 – B

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C09B 19/00 (2006.01) C09B 15/00 (2006.01) G01N 1/30 (2006.01) G01N 15/14 (2006.01) G01N 21/49 (2006.01) G01N 33/487 (2006.01) G01N 33/49 (2006.01) G01N 33/50 (2006.01) G01N 33/52 (2006.01) G01N 33/80 (2006.01)

Patent

CA 2077789

Methods for characterizing and distinguishing reticulocytes and mature red blood cells use reagent compositions which include an organic cationic dye for staining the reticulocytes in the blood sample and a buffer solution for maintaining a pH of about 6 to about 9. The dyes may be the red excitable fluorescent dye Oxazine 750, or the blue excitable fluorescent dyes Acridine Orange or derivatives of Acridine Orange. When a zwitterionic surfactant is included in the reagent composition for isovolumetric sphering of the reticulocytes and erythrocytes, and the reagent composition and whole blood sample mixture is passed through the sensing region of a flow cytometer, the light scattered through at least one angular interval and that fluoresced by each cell is measured, the erythrocytes can be distinguished from reticulocytes and the volume, hemoglobin concentration and the hemoglobin content of each reticulocyte or erythrooyte, and the mean cell volume, mean corpuscular hemoglobin concentration, and mean cell hemoglobin of the reticulocytes and/or erythrocytes are calculated from the measured cell-by-call volume and hemoglobin concentration.

es méthodes pour caractériser et distinguer les réticulocytes et les hématies adultes font appel à des composés réactifs qui comprennent un colorant organique basique pour teindre les réticulocytes de l'échantillon sanguin et une solution tampon pour maintenir un pH d'environ 6 à 9. Le colorant peut être le colorant fluorescent oxazine 750 excitable par le rouge ou le colorant fluorescent orange d'acridine excitable par le bleu ou un dérivé d'orange d'acridine. Lorsqu'un tensio-actif zwittérionique est inclus dans le composé réactif pour la sphéricité isovolumétrique des réticulocytes et des érythrocytes et que le composé réactif et le mélange global d'échantillon sanguin passent dans le capteur du cytomètre de flux, la lumière diffusée à travers au moins un intervalle angulaire et celle fluorescée par chaque globule sont mesurées, les érythrocytes peuvent être distingués des réticulocytes et le volume, la concentration d'hémoglobine et la teneur en hémoglobine de chaque réticulocyte ou érythrocyte, ainsi que le volume moyen des globules, la concentration moyenne en hémoglobine corpusculaire et la teneur moyenne en hémoglobine des réticulocytes et/ou des érythrocytes sont calculés à partir du volume et de la concentration en hémoglobine mesurés pour chaque globule.

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