Beckman Coulter, Inc.

Fabrice Malergue

Andreas Van Agthoven

Enrique Rabellino

Ravinder K Gupta

Christophe P Godefroy

Meryl A Forman

Lori A Charie

Jean-Marc Busnel

Francisco Estevez-Labori

Joseph Wolfers

Emmanuel Gautherot

Jean Marc Busnel

This disclosure is directed to methods and devices () associated with Point of Care medical testing and diagnostics. More specifically, methods and devices are described which provide a quick and streamlined way to prepare blood samples for analysis using flow cytometers, microscopes, and other analysis platforms. The benefits include a reduction in the time, resources, and expertise needed for preparing those blood samples without compromising the accuracy and efficacy of diagnosing diseases or identifying specific particulates from those blood samples.

Collection and preparation of blood samples for point-of-care diagnostics

The present invention relates to methods for labeling intracellular and extracellular targets of leukocytes, as well as to kits for performing said methods.

Method for labeling intracellular and extracellular targets of leukocytes

Systems and methods are provided for determining the age of cellular hemoglobin in individual red blood cells in a blood sample by determining the percentage of HbA1c. In embodiments, a method includes measuring side scatter and fluorescence of the individual red blood cells and identifying immature red blood cells and mature red blood cells from the side scatter and fluorescence measurement. In embodiments, data collected includes the exact number of red blood cells, the fraction limits in fluorescence and side scatter units, the mean value of each fraction in fluorescence and side scatter units, the mean FL1 values per fraction in arbitrary units and the mean side scatter values per fraction in arbitrary units. In embodiments, a method also includes deriving a HbA1c content from the measured mean fluorescence of the individual red blood cells and determining the percentage of HbA1c from the HbA1c content and the hemoglobin content of the red blood cells.

Systems and methods to determine the age of cells

A reference control for cell by cell analysis on flow cytometric analyzers contains cellular analogs made of permeated blood cells containing therein aggregated intracellular proteins and preserved antigenic sites thereof, having cellular membrane permeable to antibodies and a suspension medium. The reference control is frozen after being prepared and thawed prior to use. The cellular analogs further contain a fluorescence marker therein. Further disclosed are a method of making the reference control and a method using the reference control, as an internal or stand-alone control, for measurements of cellular hemoglobin and cellular hemoglobin variant of a blood sample.

Reference control for cell by cell analysis

Novel compositions containing (i) ligand conjugated phycobiliprotein tandem dyes and (ii) unconjugated phycobiliproteins are provided wherein the phycobiliproteins are derived from the same bacterial or eukaryotic algae species. The phycobiliproteins must be the same or different, but must contain some non-crosslinked subunits which exchange. Also provided are methods for preparing these compositions, kits containing these compositions or components of the same, and methods of using these compositions for cellular and non-cellular analysis.

Multicolor flow cytometry compositions containing unconjugated phycobiliproteins

A cell permeabilization and stabilization reagent and method of use are disclosed. The reagent contains a N-acyl sarcosine or a salt thereof, a pH adjusting agent to adjust pH of the reagent in a range from about 4 to about 6; and an aqueous medium; the reagent having a low ionic strength defined by a conductivity of less than 9.0 mS/cm. The reagent further contains bovine serum albumin and glycerol. The reagent may further include an alkyl sulfate surfactant. Upon incubating the cells with the reagent, the reagent permeates the cellular membrane to allow penetration of an intracellular marker, causes intracellular protein aggregation within the cellular membrane, while preserves a cellular constituent for binding with a cellular marker for subsequent analysis by flow cytometry.

Cell permeabilization and stabilization reagent and method of use

A method of measuring cellular hemoglobin of a blood sample includes mixing a blood sample with a permeation reagent, and incubating the sample mixture to permeate cellular membrane of red blood cells and to cause hemoglobin aggregation within the cells; adding a neutralization reagent to inhibit further reactions of the permeation reagent; performing a cell-by-cell measurement of side scatter signals of the red blood cells in the sample mixture on a flow cytometer; and obtaining cellular hemoglobin (Hgb) of each red blood cell using the obtained side scatter signals. The method further includes measuring cellular hemoglobin of reticulocytes (Hgb) by differentiating reticulocytes using a simultaneous fluorescence measurement. The method also includes measuring cellular percentage of a hemoglobin variant in mature red blood cells or reticulocytes by adding a fluorescent antibody in the neutralization reagent and detecting fluorescence signals of antibody bound hemoglobin variant.

Growing community of inventors

Marseilles, France

Fabrice Malergue