Coulter International Corp.

Beckman Coulter, Inc.

Jing Li

Yi Li

Carole Jo Young

Nery Ortiz

Ted W Britton

Min Zheng

Wayne Goldson

Ignacio Figueroa

Jiuliu Lu

Eric Malcolm Grace

Sandra Socarras

Shuliang Zhang

Maritza Lavernia

Michael J Switzer

Environmentally-friendly, aqueous concentrated reagent compositions are provided for dilution and use in suitable hematology analyzers for analyzing blood cells including for enumeration and sizing of blood cells, determination of hemoglobin parameters and differentiation of leukocyte subpopulations in a single blood cell sample.

Green concentrated reagent for hemotology systems

Green concentrated reagent for hematology systems

A method of correction of particle interference to hemoglobin measurement of a blood sample on a hematology analyzer is provided. The method includes mixing an aliquot of a blood sample with a lytic reagent to lyse red blood cells and forming a sample mixture; measuring absorbance of the sample mixture at a predetermined wavelength of a hemoglobin chromogen formed in the sample mixture, and obtaining an apparent hemoglobin concentration of the blood sample using obtained absorbance; measuring concentration and size of cellular particles remaining in the sample mixture; removing contribution of the cellular particles to the apparent hemoglobin concentration using the concentration and the size of the cellular particles to obtain a corrected hemoglobin concentration of the blood sample; and reporting the corrected hemoglobin concentration of the blood sample.

Method of correction of particle interference to hemoglobin measurement

Methods of prevention and correction of white blood cell interferences to the measurements of mean cell volume (MCV), red blood cell concentration (RBC) and hematocrit (Hct) of a blood sample are disclosed. The interference to MCV can be prevented by identifying a valley between the red blood cell and white blood cell modes on the red blood cell distribution histogram, defining a red blood cell region using the valley and calculating MCV within the defined region. Alternatively, a curve fit of the white blood cell population on the red blood cell distribution histogram can be used to exclude the white blood cells. RBC can be corrected by subtracting the white blood cell concentration obtained from the analysis of a second aliquot sample, when the white blood cell concentration (WBC) exceeds a predetermined criterion. Hct of the blood sample can be calculated using the obtained MCV and RBC.

Method of preventing white blood cell interferences to red blood cell measurements of a blood sample

Methods of using a reference control composition containing a nucleated red blood cell component for measurement of nucleated red blood cells are disclosed. The nucleated red blood cell component is made of stabilized or processed nucleated blood cells which have impedance and optical properties simulating the impedance and optical properties of human nucleated red blood cells under a blood lysing condition as measured by a specific measurement. The methods include mixing the reference control composition with a lytic reagent, and analyzing the control sample mixture on a flow cytometric instrument by impedance, impedance and optical measurement, or DC impedance and a multi-dimensional measurement of light scatter, radio frequency and a second DC impedance; and reporting the simulated nucleated red blood cells in the reference control composition.

Method of using a reference control composition for measurement of nucleated red blood cells

A method for measuring immature granulocytes in a blood sample is described. The method includes the steps of lysing red blood cells of a blood sample with a lytic reagent, analyzing the sample mixture by DC impedance measurement, determining immature granulocytes from obtained DC histogram, and reporting immature granulocytes in the blood sample. The method further includes measuring nucleated red blood cells in the blood sample.

Method for measurement of immature granulocytes

A method for differentiating and enumerating nucleated red blood cells in a blood sample is described. The method includes the steps of lysing red blood cells of a blood sample with a lytic reagent, measuring nucleated blood cells by DC impedance measurement in a non-focused flow aperture, differentiating nucleated red blood cells from other cell types, and reporting nucleated red blood cells in the blood sample. The method further includes subtracting nucleated red blood cells and other interference materials from the count of remaining blood cells, and reporting a corrected white blood cell count of the blood sample. Additionally, the method further includes measuring spectrophotometric absorbance of the sample mixture at a predetermined wavelength of a hemoglobin chromogen formed upon lysing the blood sample, and reporting hemoglobin concentration of the blood sample.

Method for measurement of nucleated red blood cells

A lytic reagent composition for measuring nucleated blood cells in a blood sample is described. The lytic reagent composition contains a quaternary ammonium surfactant, an ethoxylated phenol, and an ethoxylated alcohol. When mixed with a blood sample, the lytic reagent composition lyses red blood cells and enables a differentiation of nucleated red blood cells from other cell types by DC impedance measurement. The lytic reagent composition can further contain an organic ligand for determining total hemoglobin concentration of a blood sample photometrically. Further disclosed is a lytic reagent system including the lytic reagent composition and a diluent. In addition, a single reagent composition containing salts is also disclosed, which can be used without a separate diluent. The lytic reagent compositions can be used for concurrent measurement of nucleated red blood cells, WBC, and hemoglobin of a blood sample.

Lytic reagent composition for determination of nucleated blood cells

This invention relates to lytic reagents and methods of using the lytic reagents for automatically determining leukocyte subpopulations in blood. More specifically, the new lytic reagents selectively lyse red blood cells and certain leukocyte subpopulations, which enables the differentiation of at least one subpopulation of leukocytes. The lytic reagents contain an ethoxylated long chain amine, a quaternary ammonium salt and an acid. When used in combination with a second lytic reagent system, one is able to obtain at least a five part differential of leukocytes by impedance and light scatter measurements, by impedance and radio frequency measurements, or by radio frequency and light scatter measurements.

Composition and method for differentiation of basophils and eosinophils in blood

This invention relates to lytic reagents and methods of using the lytic reagents for automatically determining leukocyte subpopulations in blood. More specifically, the new lytic reagents selectively lyse red blood cells and certain leukocyte subpopulations, which enables the differentiation of at least one subpopulation of leukocytes. The lytic reagents contain a polyoxyethylene based surfactant, a quatemary ammonium salt and an acid. When used in combination with a second lytic reagent system, one is able to obtain at least a five part differential of leukocytes by impedance and light scatter measurements, by impedance and radio frequency measurements, or by radio frequency and light scatter measurements.

Composition and method for differentiation of basophil and eosinophil subpopulations of leukocytes in blood

A method is provided for differentiation of nucleated red blood cells. In addition, the method provides for a concurrent differentiation of leukocytes in a blood cell sample by suitable electronic and optical measurements. The method includes exposing a blood cell sample to a reagent system to lyse mature red blood cells and subsequently analyzing nucleated red blood cells in a flow cell by optical analysis. A concurrent differentiation of nucleated blood cells and leukocytes can be performed using electronic and optical analysis. The electronic and optical analysis includes light scatter and impedance measurements. This method eliminates the use of nuclear stain for identification of nucleated red blood cells. The method of the present invention, for the first time, reports differentiation and enumeration of nucleated red blood cells without using fluorescence.

Method for differentiation of nucleated red blood cells

A method is provided for differentiation of reticulocytes. In addition, the method provides for a concurrent differentiation of leukocyte subpopulations in a blood cell sample by suitable electronic and optical measurements. The method includes exposing a blood cell sample to a reagent system to lyse mature red blood cells and subsequently analyzing reticulocytes in a flow cell by optical analysis. A concurrent differentiation of reticulocytes and leukocytes can be performed using electronic and optical analysis. The electronic and optical analysis includes light scatter and impedance measurements. This method does not require the use of nuclear stain for differentiation of reticulocytes.

Growing community of inventors

Miami, FL, United States of America

Jing Li