Beckman Coulter, Inc.

Enrique Rabellino

Andreas Van Agthoven

Fabrice Malergue

Christophe P Godefroy

Carlos Aparicio

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

The present specification discloses an automated method and apparatus useful for separating and harvesting cells of interest, e.g., mononuclear cells, in a whole blood sample. The method of the invention uses the aspirating/dispensing probe of an automated sample preparation instrument to underlay a density gradient medium beneath a whole blood sample in a centrifugation tube, and the same probe is used to harvest cells of interest from a cell layer formed in the tube as a result of a centrifugation step. In harvesting cells, the probe is advanced inside the tube by a fixed, predetermined distance at which the probe tip (i.e., its aspiration port) is known to be located at, or within a predetermined distance below, the bottom of the cell layer. A predetermined volume of liquid is then aspirated through the probe tip, whereby most cells of interest (and more than 90% of those cells that can be harvested by a flawless manual method) are removed from the cell layer and collected for analysis. Preferably, the probe is caused to move laterally with respect to the tube during the aspiration of the cell layer, whereby cells offset from the center of the container are readily harvested.

Method and apparatus for separating and harvesting cells from a whole blood sample

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.

Growing community of inventors

Miami, FL, United States of America

Enrique Rabellino