Reagents, Systems and Methods for Analyzing White Blood Cells
AssigneeAbbott Laboratories
InventorJiong Wu
Life of this patent
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Abstract
Aspects of the invention include WBC analysis reagents, systems and methods that can be used for analyzing a sample of whole blood to identify, classify, and/or quantify white blood cells (WBC) and WBC sub-populations in the sample. The WBC analysis reagents of the present disclosure generally include at least one membrane-permeable fluorescent dye, a WBC protecting reagent, and a surfactant. In some embodiments, the WBC reagents include a suitable amount of an osmolality adjusting component to adjust the osmolality of the WBC reagent into a desired range.
Claims53
A method of performing a white blood cell (WBC) analysis with an automated hematology analyzer, the method comprising:
- (a) diluting a sample of whole blood with a WBC analysis reagent, wherein the WBC analysis reagent comprises:
- a membrane-permeable fluorescent dye that labels a plurality of nucleus-containing cells in the sample;
- and
- an osmolality adjustment component that separates a plurality of WBC subpopulations from one another when analyzed with the hematology analyzer;
- (b) incubating the diluted blood sample of step (a) for an incubation period of less than about 25 seconds, at a temperature ranging from about 30° C. to about 50° C.;
- (c) delivering the incubated sample from step (b) to a flow cell in the hematology analyzer;
- (d) exciting the incubated sample from step (c) with an excitation source as the incubated sample traverses the flow cell;
- (e) collecting a plurality of light scatter signals and at least one fluorescence emission signal from the excited sample;
- and
- (f) performing a WBC differential analysis based on all the signals collected in step (e), while removing from consideration any particles within the diluted blood sample that do not meet a fluorescence threshold based on the at least one fluorescence emission signal.
A system for conducting a white blood cell (WBC) differential analysis on a sample of whole blood, the system comprising:
- (a) a hematology analyzer, the hematology analyzer comprising:
- an excitation source positioned to excite particles within the blood sample;
- a plurality of detectors including (1) an axial light loss detector positioned to measure axial light loss from the excited blood sample, (2) an intermediate angle scatter detector positioned to measure intermediate angle scatters from the excited blood sample, (3) a polarized side scatter detector positioned to measure large angle polarized side scatters from the excited blood sample, (4) a depolarized side scatter detector positioned to measure large angle depolarized side scatter from the excited blood sample, and (5) a fluorescence detector positioned to measure fluorescence emitted from the excited blood sample;
- and
- a processor configured to:
- (I) receive the measurements of (1) axial light loss, (2) intermediate angle scatters, (3) large angle polarized side scatters, (4) large angle depolarized side scatters, and (5) fluorescence from the plurality of detectors, and
- (II) perform a WBC differential analysis of the blood sample, based on all five measurements, for particles that emit fluorescence above a fluorescence threshold;
- and
- (b) a reagent for analyzing WBCs in the sample, the reagent comprising:
- a membrane-permeable fluorescent dye;
- and
- an osmolality adjustment component;
- wherein the concentration of the membrane-permeable fluorescent dye is sufficient to facilitate identification of one or more cells in the sample that contain a nucleus using the hematology analyzer;
- and
- wherein the concentration of the osmolality adjustment component is sufficient to facilitate identification of a plurality of subpopulations of WBCs in the sample using the hematology analyzer.
Dependencies are inferred from the claim text.