Method for three-volume differential determination of lymphocyte, monocyte and granulocyte populations of leukocytes
First Claim
1. A method for the volumetric differentiation of leukocytes into at least two populations which are identified as monocytes and granulocytes, respectively, from a whole blood sample, using an automatic particle analyzing system, said method comprising the steps of:
- I. supplying, for use by the analyzing system for its mixing and analyzing, the whole blood sample and a volume of an isotonically balanced blood diluent; and
II. mixing the whole blood sample and the diluent by the analyzing system with a therein supplied lysing reagent in such a manner that the lysing reagent is at a significantly weak concentration and is supplied at a significantly slow rate, so as to subject the leukocytes to a low gradient of lytic shock, so as to result in the volumetric modification of the individual blood cells of at least one of the populations of leukocytes for a significant period of time to thereby enable the automatic differentiation of the leukocyte populations by the analyzing system;
the lysing reagent comprising a mixture of an aqueous solution of quaternary ammonium salts having surface acting properties.
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Abstract
A method and reagent system is described for differential determination of three-populations of leukocytes (lymphocyte, monocyte and granulocyte), using an automatic counting system. The reagent system includes a blood diluent and lysing reagent. The blood diluent is an osmotically balanced aqueous solution of ingredients at a preselected pH for maintaining erythrocyte morphology, blood cell stabilizing, buffering and bacteriostatic action. The lysing reagent is a mixture of an aqueous solution of quaternary ammonium salts, which lysing reagent is added to the diluted blood under more mild conditions of concentration and at a slower rate than is current practice in order to obtain an unexpected volumetric modification of at least one of the three-populations of leukocytes, whereby volumetric differential analysis can be accomplished.
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Citations
21 Claims
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1. A method for the volumetric differentiation of leukocytes into at least two populations which are identified as monocytes and granulocytes, respectively, from a whole blood sample, using an automatic particle analyzing system, said method comprising the steps of:
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I. supplying, for use by the analyzing system for its mixing and analyzing, the whole blood sample and a volume of an isotonically balanced blood diluent; and II. mixing the whole blood sample and the diluent by the analyzing system with a therein supplied lysing reagent in such a manner that the lysing reagent is at a significantly weak concentration and is supplied at a significantly slow rate, so as to subject the leukocytes to a low gradient of lytic shock, so as to result in the volumetric modification of the individual blood cells of at least one of the populations of leukocytes for a significant period of time to thereby enable the automatic differentiation of the leukocyte populations by the analyzing system; the lysing reagent comprising a mixture of an aqueous solution of quaternary ammonium salts having surface acting properties. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20)
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21. A method for the volumetric differentiation of at least the monocyte and granulocyte populations obtained from a whole blood sample, said method comprising the steps of:
supplying and mixing the whole blood sample and a lysing reagent, the lysing reagent is at a significantly weak concentration and is supplied at a significantly slow rate so as to result in a low gradient of lytic shock and thereby the volumetric modification of the individual blood cells of at least one of the populations of monocytes and granulocytes for a significant period of time to thereby enable the volumetric differentiation of these populations.
Specification