Beckman Coulter, Inc.

Clas Automation

Coulter International Corp.

Placidem, LLC

Carlos Aparicio

Frank J Lucas

Michael L Bell

Alexander Burshteyn

John A Maples

Jack D McNeal

Sandra Socarras

Enrique Rabellino

Oilda Rubio

Julie Wilkinson

Cecilia Smith

Nazle Kuylen

John W Joubran

Paul W Price

Maria Elena Insausti

Jeff Hrush

Adam Burke

Nicholas Crabtree

Juan Carlos Aparicio

Felipe Echeverri

Daniel Luis Granda

Fluid layering device for centrifugal tube

Blood fractionation tube

Pipette with a flared end

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 method and apparatus for preparing biological cell samples for intracellular analysis. The invention is based upon the recognition that many of the steps of the conventional methods for such sample preparation can be eliminated, leading to a process that readily lends itself to automation and the advantages associated therewith. The method of the invention comprises the steps of (a) cell-fixation, (b) permeabilization and (c) staining (or labeling) of intracellular molecules of interest by probes that are readily detectable by flow cytometric techniques, all without any intervening cell-washing (and re-suspension) steps. Rather, the single cell-washing step is effected after these three steps have been carried out. Preferably, the washing step is carried out by passing the fixed, permeabilized and stained cell sample through a semi-permeable membrane that serves to filter out (by transmission) interferants to waste while retaining the cells of interest.

Method and apparatus for preparing cell samples for intracellular antigen detection using flow cytometry

A method for utilizing a filtration device for removing interferants from a test sample containing a mixture of a composition of interest and interferants in an automated apparatus is disclosed. The filtration device includes a microporous hollow fiber membrane having a plurality of pores sized to retain the composition of interest while allowing smaller diameter interferants to pass through the membrane.

Apparatus and method for sample purification and analysis

Growing community of inventors

Miami, FL, United States of America

Carlos Aparicio