Abbott Laboratories Corporation

Young Ran Kim

Johanna Kantor

Roderick W Larue

Peter E Lobban

Vernon L Chupp

John Paul Stuart

Stephen D Stroupe

Suresh N Mehta

Michael W Yee

James E Gill

Jean Emiko Kihara

Josefino C Sagala

Sue E Luptovic

Kenneth E Iles

Willie J Cowart

Ronny Van't Oever

James C Bearden, Jr

Marilou Z Landayan

A method for distinguishing erythroblasts from bacteria by automated hematology analyzers, such as, for example, the CELL-DYN&#xae; 4000 automated hematology analyzer and the CELL-DYN&#xae; Sapphire&#x2122; automated hematology analyzer. Bacterial cells scatter light and fluoresce differently than do red blood cells, white blood cells, erythroblast nuclei, and platelets. Signals generated by bacteria are distinguishable from those of erythroblasts because the signals generated by erythroblast nuclei are sufficiently unique that erythroblast nuclei can be distinguished from signals generated by bacteria. Signals generated by platelets, lysed red blood cell ghosts, and other cell debris are blocked by the triple-trigger circuitry of the hematology analyzer, because all of the signals generated by noise are below the AND/OR thresholds. Algorithm(s) in the software of the system detect and count signals generated by bacteria by means of the location and the shape of the signals generated by bacteria and calculate the concentration of bacteria per unit of body fluid. In addition, certain body fluids, such as, for example, synovial fluid, can be pretreated with a viscosity reducing agent for a short period of time to reduce the viscosity of the body fluid prior to analyzing a sample of the body fluid by an automated hematology analyzer.

Method for classifying and counting bacteria in body fluids

Methods and apparatus are disclosed for determining the volume, hemoglobin concentration, maturity and cell shape of mammalian red blood cells in a whole blood sample and simultaneously monitoring system standardization. Methods for distinguishing red blood cells from other cellular particles, prior to the red blood cell analysis are also disclosed. Red blood cells are passed through a beam of light in single file at a selected wavelength, obtaining an initial cytogram by means of the resultant magnitude of one light loss signal and one forward angle light scatter signal at a selected angular interval and a third side angle light scatter or two forward angle light scatter signals at a selected angular intervals and a third side-angle light scatter signal, projecting the cytogram, point by point, onto a pre-calibrated 3-dimensional surface containing grid lines of volume and hemoglobin concentration and determining accurate values of cell volume and hemoglobin concentration.

Optical method and apparatus for red blood cell differentiation on a cell-by-cell basis, and simultaneous analysis of white blood cell differentiation

Hematology reference control cells and method of manufacture. The invention relates to the methods of preparing stable white blood cell (&ldquo;WBC&rdquo;) and nucleated red blood cell (&ldquo;NRBC&rdquo;) fractions and the hematology control reagents containing such stablized cells for primary use on a multi-angle light scatter based hematology instrument.

Hematology reference control and method of preparation

A device for analyzing a whole blood sample is provided. The device comprises a conventionl hematology analyzer integrated with a fluorescence cyometry analyzer. A controller is provided for controlling the analyzers, obtaining and utilizing data from both and reporting a quantitative result. Methods are also provided for analyzing a whole blood sample. One such method comprises the steps of performing on a single instrument an analysis of impedance associated with the blood sample, an analysis of light scatter associated with the blood sample, and an analysis of fluorescence associated with the blood sample. Data is collected and utilized. A result is reported.

Method and apparatus for performing automated analysis

A method for the simultaneous and quantitative, flow cytometric analysis of nucleated red blood cells (NRBC), white blood cells (WBC), damaged white blood cells and a white blood cell subclass differential (WBC/Diff) is provided. The method includes mixing an aliquot of a whole blood sample with a reagent system comprising a red blood cell (RBC) lysing component which lyses RBCs and NRBCs while minimizing damage to WBC cellular membranes and a membrane-impermeant nucleic acid stain which stains exposed NRBC nuclei and damaged WBCs, subjecting the stained aliquot to flow cytometric light measurements, obtaining at least one signal for the parameters of fluorescence (FL) and scattered light at a first and a second range of scatter angles, qualifying the obtained signals using AND/OR logic wherein to be qualified, a signal must be greater than the second scatter signal threshold AND also greater than either the first scatter signal threshold OR the FL threshold, constructing a three-dimensional plot of qualified intensity signals of fluorescence and scattered light from the detected signals, and differentiating the NRBC, WBC, damaged WBC and WBC/Diff from the constructed three-dimensional plot and determining the number of cells of each.

Method for simultaneous analysis of cell viability, nucleated red blood

A cyanide-free method and reagent for determining the concentration of total hemoglobin in a whole blood sample accurately in less than 10 seconds including a ligand selected from the group consisting of imidiazole, imidazole derivatives, N-hydroxyacetamide, N-hydroxyl amine, pyridine, oxazole, thiazole, pyrazole, pyrimidine, purine, quinoline, and isoquinoline, and a surfactant with strong erythrolytic capability selected from the group consisting of lauryl dimethylamine oxide and octylphenoxy polyethoxyethanol. The reagent pH is adjusted to about 11 to about 14. Rapid mixing of the reagent with a blood sample leads to the formation of a stable chromogen whose absorbance can be measured between 540 and 550 nm. The cyanide-free reagent is ideal for use on an automated high through-put clinical hematology analyzer.

Cyanide-free reagent and method for the determination of hemoglobin

Hematology reference control cells and method of manufacture. The invention relates to the methods of preparing stable white blood cell ('WBC') and nucleated red blood cell ('NRBC') fractions and the hematology control reagents containing such stablized cells for primary use on a multi-angle light scatter based hematology instrument.

A method and reagent for the simultaneous or independent enumeration of reticulocytes in a whole blood sample, without the need to separately incubate the sample and reagent. The reagent contains a reticulocyte staining amount of an unsymmetrical cyanine dye, from about 40 mM to about 60 mM of a buffer selected from the group consisting of imidazole, Tris and Bis-Tris and a dye stabilizing amount of a non-ionic surfactant selected from the group consisting of N, N-bis�3-D-Glucon-amidopropyl! cholamide and a polyoxypropylene-polyoxyethylene block copolymer. The reagent has a pH from about 6.8 to about 7.2 and an osmolarity adjusted to about 280 to about 310 mosm/l with a mono-, or di-, valent alkali salt selected from the group consisting of NaCl, KCl, LiCl, CaCl.sub.2, MgCl.sub.2 and ZnCl.sub.2. The method utilizes the reagent in a no incubation process that also allows for the simultaneous determination of CBC as well as reticulocyte counts and maturity indices.

Methods for the rapid analysis of the reticulocytes

A method and reagent for the simultaneous or independent enumeration of reticulocytes in a whole blood sample, without the need to separately incubate the sample and reagent. The reagent contains a reticulocyte staining amount of an unsymmetrical cyanine dye, from about 40 mM to about 60 mM of a buffer selected from the group consisting of imidazole, Tris and Bis-Tris and a dye stabilizing amount of a non-ionic surfactant selected from the group consisting of N, N-bis�3-D-Glucon-amidopropyl! cholamide and a polyoxypropylene-polyoxyethylene block copolymer. The reagent has a pH from about 6.8 to about 7.2 and an osmolarity adjusted to about 280 to about 310 mOsm/l with a mono-, or di-, valent alkali salt selected from the group consisting of NaCl, KCl, LiCl, CaCl.sub.2, MgCl.sub.2 and ZnCl.sub.2. The method utilizes the reagent in a no incubation process that also allows for the simultaneous determination of CBC as well as reticulocyte counts and maturity indices.

Compositions and methods for the rapid analysis of reticulocytes

Method for performing automated hematology and cytometry analysis

A multipurpose reagent system for rapid analysis of a whole blood sample allowing the determination of at least five classes of peripheral white blood cells, nucleated red blood cells, and lymphocyte immunophenotyping on automated hematology instrumentation. The multipurpose reagent system lyses red cells rapidly, while it concurrently fixes white cells and preserves surface antigens on lymphocytes. The multipurpose reagent system comprises from about 3 to 7 grams per liter of a non-quaternary ammonium salt, from about 0.04 to about 0.10 percent by volume of an aliphatic aldehyde with one to four carbons, from about 10 mM to about 20 mM of a non-phosphate buffer which is inert to the aliphatic aldehyde, and a sufficient amount of water to give a pH between 5.5 and 7.5 and an osmolality of between about 160 to about 310 mOsm per liter.

Method of using a multi-purpose beagent for subclassification of

A device for analyzing a whole blood sample is provided. The device comprises a conventional hematology analyzer integrated with a fluorescence cytometry analyzer. A controller is provided for controlling the analyzers, obtaining and utilizing data from both and reporting a quantitative result. Methods are also provided for analyzing a whole blood sample. One such method comprises the steps of performing on a single instrument an analysis of impedance associated with the blood sample, an analysis of light scatter associated with the blood sample, and an analysis of fluorescence associated with the blood sample. Data is collected and utilized. A result is reported.

A cyanide-free method and reagent for determining the concentration of total hemoglobin in a whole blood sample accurately in less than 10 seconds comprising a ligand selected from the group consisting of imidiazole, imidazole derivatives, N-hydroxyacetamide, N-hydroxyl amine, pyridine, oxazole, thiazole, pyrazole, pyrimidine, purine, quinoline, and isoquinoline, and a surfactant with strong erythrolytic capability selected from the group consisting of lauryl dimethylamine oxide and octylphenoxy polyethoxyethanol. The reagent pH is adjusted to about 11 to about 14. Rapid mixing of the reagent with a blood sample leads to the formation of a stable chromogen whose absorbance can be measured between 540 and 550 nm. The cyanide-free reagent is ideal for use on an automated high through-put clinical hematology analyzer.

Embodiments described herein provide apparatuses and methods for preparing fluid. In one method, a heater thermally associated with a housing for receiving fluid is operated to apply thermal energy to the housing. A second predetermined volume of a second fluid is introduced to the housing. The second predetermined volume of the second fluid is moved with the housing for a first predetermined time period. The first predetermined volume of the first fluid is introduced to the housing. The first predetermined volume of first fluid and the second predetermined volume of second fluid are moved with the housing for a second predetermined time period. The first predetermined volume of first fluid and the second predetermined volume of second fluid are removed from the housing. Another embodiment provides an apparatus comprising a first housing for containing fluid. A second housing is operatively connected with the first housing. A prime mover is fixedly-attached to the second housing and is operatively connected with the first housing such that the first housing moves responsive to the prime mover and such that the second housing remains substantially stationary with respect to the prime mover. A fluid inlet is fluidly connected with the first housing for supplying fluid to the first housing. A fluid outlet is fluidly connected with the first housing for removing fluid from the first housing.

Cell suspension preparation apparatus and method

A method and a device for the simultaneous and quantitative, flow cytometric analysis of nucleated red blood cells (NRBC) and white blood cells (WBC) in a whole blood sample. The method includes the lysis of red blood cells (RBC) and NRBC cytoplasm from an aliquot of a whole blood sample to eliminate RBC and to expose the NRBC to a vital nuclear stain and the inhibition of the permeation of the nuclear stain into the WBC, subjecting the stained aliquot to flow cytometric light measurements, obtaining at least one signal for parameters including scattered light at a first and a second range of scatter angles and fluorescence (FL), qualifying the obtained signals by using the combination logic wherein a qualified signal must be greater than the second scatter signal threshold, while at the same time it must be greater than either the first scatter signal threshold or the FL threshold {[(first scatter angle signal OR FL signals) AND second scatter angle signal]}, constructing a three-dimensional plot of qualified intensity signals of fluorescence and scattered light from the detected signals, and differentiating the NRBC and WBC from the constructed three-dimensional plot and determining the number of cells of each.

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Sunnyvale, CA, United States of America

Young Ran Kim