Becton Dickinson and Company Limited

Hematologics, Inc.

Hermatologics, Inc.

Michael R Loken

Leon W Terstappen

Sanjaya N Joshi

Stephen C Wardlaw

Robert A Levine

Rodolfo R Rodriguez

Diether J Recktenwald

Virendra O Shah

Chia H Chen

Rickie S Kerndt

Andrew P Voigt

Curt I Civin

Fridtjof Lund-Johansen

Ronald A Mickaels

Johanna Olweus

Philip R Greipp

Nick J Gonchoroff

Elizabeth L Evans

David W Houck

Jerry A Katzmann

Robert A Kyle

Richard K Dost

Shiang Huang

A normal set of cells is characterized using flow cytometry. A centroid and radius are defined for a set of clusters in an n-dimensional space corresponding to a normal maturation for a cell lineage in the normal set of cells. A test set of cells is characterized using flow cytometry and the characterization is compared to the defined set of clusters. Support Vector Machine (SVM) subroutines are employed to identify reference populations of interest by generating multidimensional boundary definitions. These boundary definitions may be used to identify reference populations to use in defining or refining a centroid line or a radius or radii defining a set of normal clusters, and to characterize and compare a test set of cells to the defined set of normal clusters.

System, method, and article for detecting abnormal cells using multi-dimensional analysis

A system, method, and article for diagnosing a test set of biological cells. For example, in one embodiment a normal set of cells is characterized using flow cytometry. A centroid and radius are defined for a set of clusters in an n-dimensional space corresponding to a normal maturation for a cell lineage in the normal set of cells. A test set of cells is characterized using flow cytometry and the characterization is compared to the set of clusters.

The present invention provides improved methods for collecting cells for subsequent analysis. More specifically, the invention provides methods for collecting purified cell samples from a flow cytometer onto a solid support on which cell lysis occurs and genetic material is immobilized.

Method for collecting purified cells

One or more population of cells enriched for human hematopoietic stem cells is disclosed. HSC in this population of cells are capable of limited self-renewal and are capable of differentiating into all elements of the hematopoietic system. This population of cells has the phenotype of CD34.sup.+ /CD38.sup.- and more preferably CD34.sup.+ /CD38.sup.- /HLA-DR.sup.+. Cells within this population have been found to express CD13, CD33 and CD71. Hematopoietic stem cells can be used in a number of therapies, including autologous transplantation and in gene therapy.

Phenotypic characterization of the hematopoietic stem cell

A method for the non-linear, non-logrithmic transformation of one or more parameters of data gathered by means of flow cytometry is disclosed wherein the method seeks to maximize the resolution between populations of interest for each parameter while maintaining the dynamic range of the data recorded.

Method for data transformation

A method and kit for identification of the lineages and stages of hematopoietic cells in a sample wherein the cells are treated with labeled monoclonal antibodies selected to distinguish between each lineage and maturational stages within each lineage.

Method to determine the composition of bone marrow samples

A method is disclosed for the discrimination between intact and damaged cells in a sample, and more particularly, the method uses a vital nucleic acid dye to selectively stain intact versus damaged cells which then may be counted and sorted by flow cytometry means. The method also may be used in conjunction with fluorescently labelled monoclonal antibodies to simultaneously identify cellular antigens.

Method for discriminating between intact and damaged cells in a sample

A method for multi-parameter analysis of cells in a body fluid sample is described which makes use of a plurality of fluorescence measurements, comprising at least two nucleic acid dyes and at least one fluorescently labelled cell surface marker, and a plurality of light scattering measurements. A kit containing the nucleic acid dyes and cell surface marker also is described.

Method for analysis of cellular components of a fluid

In order to produce a more well defined interface between adjacent cell layers in a centrifuged sample of anticoagulated whole blood, a material which will bond one group of cells together is added to the blood sample prior to centrifugation. The bonding material must produce a high strength bond between one group of cells, but not effect the other cell types. The material is added prior to centrifugation of the blood sample.

Method for increasing agglutination of groups of cells to produce

A method is described for the detection of halodeoxyuridines taken up by cells during DNA synthesis by the use of monoclonal antibodies. The described method also allows the simultaneous detection of cell surface receptors.

Detection of cellular DNA

A method for calibrating an instrument for using that instrument to obtain at least one light-related signal from particles under analysis comprises directing an incident beam of light at calibration particles having one or more known characteristics related to the particles expected to be analyzed. Both a light signal and a noise signal from the calibration particles are detected. A measurement is made of the ratio of the detected light signal to the detected noise signal, and that measurement is reported. The measured ratio is then compared to a predetermined ratio which represents a threshold for minimum instrument performance. This method further includes adjusting, if the predetermined ratio has not been attained, the operation of the instrument, while the calibration particles are within the incident beam of light, until the measured ratio reaches the predetermined ratio whereby the instrument is calibrated for subsequently obtaining the light signal from particles to be analyzed. A kit of reagents for use in calibrating an instrument for performing two-color analysis of particles and particle standars for calibrating fluorescence analysis instruments are also part of the instant invention.

Method and materials for calibrating flow cytometers and other analysis

A method for calibrating an instrument for using that instrument to obtain at least one light-related signal from particles under analysis comprises directing an incident beam of light at calibration particles having one or more known characteristics related to the particles expected to be analyzed. Both a light signal and a noise signal from the calibration particles are detected. A measurement is made of the ratio of the detected light signal to the detected noise signal, and that measurement is reported. The measured ratio is then compared to a predetermined ratio which represents a threshold for minimum instrument performance. This method further includes adjusting, if the predetermined ratio has not been attained, the operation of the instrument, while the calibration particles are within the incident beam of light, until the measured ratio reaches the predetermined ratio whereby the instrument is calibrated for subsequently obtaining the light signal from particles to be analyzed. A kit of reagents for use in calibrating an instrument for performing two-color analysis of particles and particle standards for calibrating fluorescence analysis instruments are also part of the instant invention.

Growing community of inventors

Mercer Island, WA, United States of America

Michael R Loken