Multi-purpose blood diluent and lysing agent for differential determination of lymphoid-myeloid population of leukocytes
First Claim
1. A stromatolysing reagent for use in the determination of at least two leukocyte populations in blood, two such populations being lymphoid and myeloid, the blood being processed through a blood cell analyzer using the Coulter Principle of operation, which employs a leukocyte sensing zone, the blood having been first diluted with an electrically conductive isotonically balanced diluent, wherein said stromatolysing reagent comprises an aqueous solution of at least two quaternary ammonium salts having surface active properties and an additive which is at least one non-cationic surfactant, said salts and additive being present in sufficient amounts for positioning said leukocyte populations relative to one another and relative to a blood cell volume reference point, within the time constraints of said blood cell analyzer.
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Abstract
A method and reagent system is described for differential determination of more that one class of leukocyte populations, using an automatic blood cell analyzer. The reagent system includes a blood diluent and stromatolyzing reagent. The blood diluent is an osmotically balanced aqueous solution of ingredients for maintaining erythrocyte morphology, blood cell stabilizing, buffering and bacteriostatic action. The stromatolyzing reagent is a mixture of an aqueous solution of quaternary ammonium salts, a chromagen forming agent and an additive which is a non-cationic surfactant. The quaternary ammonium salts, with the additive, serve the purpose of positioning, relative to one another and a volume reference point, the lymphoid and myeloid populations. The additive also acts to prevent or to reduce the amount of protein deposit in the sensing orifices of the analyzer, which deposit tends to accumulate from the cell debris resulting from blood cell stromatolyzation.
102 Citations
24 Claims
- 1. A stromatolysing reagent for use in the determination of at least two leukocyte populations in blood, two such populations being lymphoid and myeloid, the blood being processed through a blood cell analyzer using the Coulter Principle of operation, which employs a leukocyte sensing zone, the blood having been first diluted with an electrically conductive isotonically balanced diluent, wherein said stromatolysing reagent comprises an aqueous solution of at least two quaternary ammonium salts having surface active properties and an additive which is at least one non-cationic surfactant, said salts and additive being present in sufficient amounts for positioning said leukocyte populations relative to one another and relative to a blood cell volume reference point, within the time constraints of said blood cell analyzer.
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18. In a method for determination of at least two leukocyte populations in blood which is being processed through a blood cell analyzer using the Coulter Principle of operation which includes the step of:
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I. diluting a blood sample with an electronically conductive isotonically balanced diluent, the improvement which comprises the step of; II. stromatolysing with a reagent comprising an aqueous solution of at least two quaternary ammonium salts having surface active properties, and an additive which contains at least one non-cationic surfactant, said salts and additive being present in sufficient amounts for affecting the normal leukocyte volume distributions during said stromatolysation so as to cause the volume distribution measurements of said blood cell analyzer to lie within predetermined limits, with reference to at least one volume reference point. - View Dependent Claims (19, 20, 21, 22)
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- 23. In a method for the size differentiation of at least two populations of leukocytes in blood, the blood being processed through a blood cell analyzer using the Coulter Principle of operation which employs a leukocyte sensing zone, the blood having been first diluted with an electrically conductive isotonically balanced diluent, the improvement wherein said stromatolysing reagent comprises an aqueous solution of at least two quaternary ammonium salts having surface active properties and an additive which is at least one non-cationic surfactant, said salts and additive being present in sufficient amounts for positioning said leukocyte populations relative to one another and relative to a blood cell volume reference point, within the time constraints of said blood cell analyzer.
Specification