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

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

G - Physics

01

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G01N 33/52 (2006.01) G01N 1/30 (2006.01) G01N 21/49 (2006.01) G01N 33/487 (2006.01) G01N 33/49 (2006.01) G01N 33/50 (2006.01) G01N 33/80 (2006.01)

Patent

CA 2077788

Methods for characterizing and distinguishing reticulocytes and mature red blood cells use reagent compositions which include organic cationic dyes for staining the reticulocytes in the blood sample and buffer solutions for maintaining a pH of about 6 to about 9. The dyes may be the blue absorption dyes Oxazine 750 or New Methylene Blue. 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 and absorbed 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 erythrocyte, and the mean cell volume, mean corpuscular hemoglobin concentration, and mean cell hemoglobin of the reticulocytes or erythrocytes are calculated from the measured cell-by-cell volume and hemoglobin concentration. One method also includes adjusting the measured absorption signals for pseudo-absorption.

Méthodes permettant de caractériser et de distinguer les réticulocytes et les érythrocytes mûrs au moyen de compositions réactives qui comprennent des teintures cationiques organiques pour colorer les réticulocytes dans l'échantillon sanguin et les solutions tampons afin de maintenir un pH d'environ 6 à 9. Les teintures peuvent être les teintures absorbantes bleues Oxazine 750 ou le nouveau bleu de méthylène. Lorsqu'un surfactant zwittérionique est inclus dans la composition réactive à des fins de sphering isovolumétrique des réticulocytes et des érythrocytes, et lorsque le mélange composition réactive et échantillon de sang total est passé à travers la zone de détection d'un cytomètre de flux, la lumière diffusée et absorbée par chaque globule est mesurée, les érythrocytes peuvent être distingués des réticulocytes et le volume, la concentration en hémoglobine et la teneur en hémoglobine de chaque réticulocyte ou érythrocyte, ainsi que le volume globulaire moyen, la concentration corpusculaire moyenne en hémoglobine, et la teneur globulaire moyenne en hémoglobine des cellules des réticulocytes ou des érythrocytes sont calculés à partir du volume et de la concentration en hémoglobine mesurées globule par globule. Une méthode consiste également à ajuster les signaux d'absorption mesurés afin de tenir compte de la pseudo-absorption.

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