Technicon Instruments Corporation

Mount Sinai School of Medicine

Miles Inc.

Bayer Corporation

Other

Siemens Healthcare Diagnostics Gmbh

Leonard Ornstein

Gena Fischer

Gregory M Colella

Sophie S Fan

Daniel Ben-David

Albert Cupo

Young R Kim

Grace E Martin

Michael J Malin

Julius Intraub

Hazel Elizabeth Williams

Bruce J Oberhardt

David Zelmanovic

Abraham Gordon

Martin J Lee

Jolanta Kunicka

Robert J Kanter

Henry Cook Waters, Iii

Pamela Elsins

Hudson R Ansley

Stanford L Adler, Sr

The present invention describes semi- and fully-automated methods and reagents therefor for the assay and analysis of body fluid samples, particularly non-blood samples. The methods and reagents are especially useful for the assay and analysis of cerebrospinal fluid (CSF) samples. The reagent compositions sphere and fix all cells in the sample in suspension. Reported results can include red blood cell (RBC) and white blood cell (WBC) counts, WBC differential values, cell-by-cell volumes and dry-mass concentrations.

Automated method and reagent therefor assaying body fluid samples such as cerebrospinal fluid (CSF)

The invention provides a bicarbonate-containing, crosslinker-free reagent composition which reverses and/or reduces sample handling effects, such as aeration-induced shrinkage and storage-induced swelling of cells in a whole blood sample, particularly for a blood sample requiring repeated sampling and/or subjected to long-term storage. The reagent composition serves as a diluent medium for a blood sample, or an aliquot or portion thereof. The pH of the reagent is in the range of about 7.2 to about 7.5, preferably 7.3. The stability of the pH 7.2-7.5 bicarbonate-containing reagent over time is maintained by storing the reagent in a flexible collapsible container which is impermeable to carbon dioxide and is comprised of a multilayered flexible material, preferably plastic, and attachable to a hematology analyzer.

METHOD AND LONG-TERM STABLE BICARBONATE-CONTAINING DILUENT COMPOSITION, AND STORAGE MEANS THEREFOR, FOR REDUCING OR REVERSING AERATION INDUCED CELL SHRINKAGE AND STORAGE INDUCED CELL SWELLING OF A WHOLE BLOOD SAMPLE

Methods for blood sample preparation using reagent compositions which include a zwitterionic surfactant for sphering of cells to eliminate orientational noise. When the reagent composition including a dye for staining a subset of cells is reacted with a blood sample, and the reaction mixture is passed through the sensing region of a flow cytometer, the light scattered and absorbed by each cell is measured, the stained cells can be distinguished from the unstained cells, and when the cells are reticulocytes and mature erythrocytes, respectively, the volume and hemoglobin concentration of each reticulocyte or erythrocyte, and the hemoglobin content, mean cell volume, mean corpuscular hemoglobin concentration, and mean cell hemoglobin of the reticulocytes or erythrocytes are calculated from the measured cell-by-cell volume and hemoglobin concentration.

Reagent compositions for use in sphering cells

An unsupervised back propagation method for training neural networks. For a set of inputs, target outputs are assigned l's and O's randomly or arbitrarily for a small number of outputs. The learning process is initiated and the convergence of outputs towards targets is monitored. At intervals, the learning is paused, and the values for those targets for the outputs which are converging at a less than average rate, are changed (e.g., 0.fwdarw.1, or 1.fwdarw.0), and the learning is then resumed with the new targets. The process is continuously iterated and the outputs converge on a stable classification, thereby providing unsupervised back propagation. In a further embodiment, samples classified with the trained network may serve as the training sets for additional subdivisions to grow additional layers of a hierarchical classification tree which converges to indivisible branch tips. After training is completed, such a tree may be used to classify new unlabelled samples with high efficiency. In yet another embodiment, the unsupervised back propagation method of the present invention may be adapted to classify fuzzy sets.

Unsupervised neural network classification with back propagation

An unsupervised back propagation method for training neural networks. For a set of inputs, target outputs are assigned 1's and 0's randomly or arbitrarily for a small number of outputs. The learning process is initiated and the convergence of outputs towards targets is monitored. At intervals, the learning is paused, and the values for those targets for the outputs which are converging at a less than average rate, are changed (e.g., 0.fwdarw.1, or 1.fwdarw.0), and the learning is then resumed with the new targets. The process is continuously iterated and the outputs converge on a stable classification, thereby providing unsupervised back propagation. In a further embodiment, samples classified with the trained network may serve as the training sets for additional subdivisions to grow additional layers of a hierarchical classification tree which converges to indivisible branch tips. After training is completed, such a tree may be used to classify new unlabelled samples with high efficiency. In yet another embodiment, the unsupervised back propagation method of the present invention may be adapted to classify fuzzy sets.

Method for unsupervised neural network classification with back

A composition is disclosed for transferring and adhering tape-mounted hisogical sections, to specimen mounting surfaces, usually glass microscope slides to expedite and simplify the safe removal of the mounting tape and the subsequent processing of the slide-mounted section. The composition is a curable polymeric mixture which has high tack prior to curing, is substantially non-diffusable and non-flowable into tissue sections and, after curing, has a refractive index substantially similar to that of the specimen section and is non-labile to conventional histological solvents and stains. A preferred composition comprises diacrylate-terminated polyurethane, a diacrylate ester of an epoxy resin and a diethoxyacetophenone initiator. It is usually formed onto the mounting surface as a solution in a conventional organic solvent, such as isopropanol or toluene.

Specimen mounting adhesive composition

A reagent composition which includes organic cationic dyes for staining the reticulocytes in a blood sample and buffer solutions for maintaining a pH of about 6 to about 9 is provided. The dyes may be the blue absorption dyes Oxazine 750 or New Methylene Blue. A zwitterionic surfactant is included in the reagent composition for isovolumetric sphering of the reticulocytes and erythrocytes. The reagent composition and whole blood sample mixture are passed through the sensing region of a flow cytometer. The light scattered and absorbed by each cell is measured; the erythrocytes can be distinguished from reticulocytes and the volume, hemoglobin concentration and the hemoglobin content of each reticulocyte or erythrocyte, and the mean cell volume, mean corpuscular hemoglobin concentration, and mean cell hemoglobin of the reticulocytes or erythrocytes are calculated from the measured cell-by-cell volume and hemoglobin concentration.

Reagent compositions for use in the identification and characterization

A reagent composition which include an organic cationic dye for staining the reticulocytes in the blood sample and a buffer solution for maintaining a pH of about 6 to about 9 is provided. The dyes may be the red excitable fluorescent dye Oxazine 750, or the blue excitable fluorescent dyes Acridine Orange or derivatives of Acridine Orange. A zwitterionic surfactant is included in the reagent composition for isovolumetric sphering of the reticulocytes and erythrocytes. The reagent composition and whole blood sample mixture is passed through the sensing region of a flow cytometer, the light scattered through at least one angular interval and that fluoresced by each cell is measured, the erythrocytes can be distinguished from reticulocytes and the volume, hemoglobin concentration and the hemoglobin content of each reticulocyte or erythrocyte, and the mean cell volume, mean corpuscular hemoglobin concentration, and mean cell hemoglobin of the reticulocytes and/or erythrocytes are calculated from the measured cell-by-cell volume and hemoglobin concentration.

Reagent compositions and their use in the identification and

Methods for characterizing and distinguishing reticulocytes and mature red blood cells use reagent compositions which include an organic cationic dye for staining the reticulocytes in the blood sample and a buffer solution for maintaining a pH of about 6 to about 9. The dyes may be the red excitable fluorescent dye Oxazine 750, or the blue excitable fluorescent dyes Acridine Orange or derivatives of Acridine Orange. When a zwitterionic surfactant is included in the reagent composition for isovolumetric sphering of the reticulocytes and erythrocytes, and the reagent composition and whole blood sample mixture is passed through the sensing region of a flow cytometer, the light scattered through at least one angular interval and that fluoresced by each cell is measured, the erythrocytes can be distinguished from reticulocytes and the volume, hemoglobin concentration and the hemoglobin content of each reticulocyte or erythrocyte, and the mean cell volume, mean corpuscular hemoglobin concentration, and mean cell hemoglobin of the reticulocytes and/or erythrocytes are calculated from the measured cell-by-cell volume and hemoglobin concentration.

Methods for the identification and characterization of reticulocytes in

Methods for characterizing and distinguishing reticulocytes and mature red blood cells use reagent compositions which include organic cationic dyes for staining the reticulocytes in the blood sample and buffer solutions for maintaining a pH of about 6 to about 9. The dyes may be the blue absorption dyes Oxazine 750 or New Methylene Blue. When a zwitterionic surfactant is included in the reagent composition for isovolumetric sphering of the reticulocytes and erythrocytes, and the reagent composition and whole blood sample mixture is passed through the sensing region of a flow cytometer, the light scattered and absorbed by each cell is measured, the erythrocytes can be distinguished from reticulocytes and the volume, hemoglobin concentration and the hemoglobin content of each reticulocyte or erythrocyte, and the mean cell volume, mean corpuscular hemoglobin concentration, and mean cell hemoglobin of the reticulocytes or erythrocytes are calculated from the measured cell-by-cell volume and hemoglobin concentration. One method also includes adjusting the measured absorption signals for pseudo-absorption.

Methods for blood sample preparation using reagent compositions which include a zwitterionic surfactant for sphering of cells to eliminate orientational noise. When the reagent composition including a for staining a subset of cells is reacted with an anticoagulated blood sample, and the reaction mixture is passed through the sensing region of a flow cytometer, the light scattered and absorbed by each cell is measured, the stained cells can be distinguished from the unstained cells, and when the cells are reticulocytes and mature erythrocytes, respectively, the volume and hemoglobin concentration of each reticulocyte or erythrocyte, and the hemoglobin content, mean cell volume, mean corpuscular hemoglobin concentration, and mean cell hemoglobin of the reticulocytes or erythrocytes are calculated from the measured cell-by-cell volume and hemoglobin concentration.

Reagent compositions and their use in sphering cells

A flow cytometry method for reproducibly detecting and counting a lymphocyte population of interest in a leukocyte suspension or whole blood sample in which the red cells are subsequently lysed. The suspension (or sample) is combined with a reagent comprising a primary antibody, either native, carrying an attached enzyme or biotin or other label, and a fixative reagent, in either order. Where the enzyme is not attached, an enzyme is coupled specifically to the primary antibody. The fixed suspension is reacted with a color-producing enzyme-cytochemical reagent. The suspension, now including stained and unstained fixed cells, is passed through a flow cytometer and the cells are characterized and counted on the basis of their light-scattering and light-absorbing properties.

Method for discriminating surface stained lymphocytes

A composition is disclosed of blood serum sample preparation for improved, more accurate and precise, electrooptical method for measuring erythrocyte volumes, individually and as an average.

Reagent mixture and composition for treating red blood cells to effect

A rapid one step, one reagent method is disclosed for the isovolumetric sphering and fixing of whole blood erythrocytes.

One step method for sphering and fixing whole blood erythrocytes

A method is described for transferring a thin tissue section affixed to a substrate by a first pressure-sensitive adhesive layer to a microscope slide which supports a second pressure-sensitive adhesive layer of polymerizable material. The tissue section is contacted to the second adhesive layer to form a laminate. Following polymerization of the second layer to form a positive bond between the tissue section and microscope slide, the substrate and first adhesive layer are completely removed to expose the tissue section, now affixed to the microscope slide by the polymerized second adhesive layer, for subsequent processing.

Growing community of inventors

White Plains, NY, United States of America

Leonard Ornstein