Features | Partner Sites | Information | LinkXpress
Sign In
FOCUS DIAGNOSTICS, INC.
AB Sciex
RANDOX LABORATORIES

Flow Cytometry Evaluated for Cord Blood Differentials

By Labmedica International staff writers
Posted on 20 Feb 2013
The potential of flow cytometry to analyze and to report leukocyte differentials on cord blood to be used for hematopoietic stem cells has been evaluated.

Currently, the reference method for white blood cell (WBC) is the microscopic manual count, but international standards advise that potential donor blood is tested for total nucleated cell count, nucleated red blood cell count (nRBCs), total number of CD34 cells, viability or potency as measured by viable CD34 cells. Colony-forming unit assays, microbial cultures, and finally a complete blood count with WBC differential are also advisable.

Hematologists at the Centre Hospitalier Universitaire (CHU; Rennes, France) analyzed 161 cord bloods between November 2010 and February 2011. WBC differentials were determined for each sample, by a cell counter, a manual method, and the flow cytometry using an antibody cocktail. WBC differential includes the count of neutrophils, eosinophils, basophils, lymphocytes, monocytes, immature granulocytes, and blasts cells.

All samples were analyzed on the HematoFlow platform (Beckman Coulter, Miami, FL, USA) within 12 hours of collection. This platform is built around Beckman Coulter's FP1000 cell preparator, along with their FC500 five-color flow cytometer. The antibody cocktail used was another Beckman Coulter product called the CytoDiff.

A good correlation was obtained for neutrophils, lymphocytes, and immature granulocytes when the manual method was compared to the flow cytometry, but was weaker for eosinophils and even lower for monocytes and basophils. The flow cytometry method employed allowed the identification of 12 cell
subtypes (neutrophils, eosinophils, basophils, B lymphocytes, T-non-cytotoxic and T cytotoxic/Natural Killer (NK) lymphocytes, CD16 negative monocytes and CD16 positive monocytes, immature granulocytes, B blasts, T blasts and non-B-non-T blasts) within a single panel of antibodies and a minimal volume of 50 µL of blood.

The authors concluded that the flow differential has some drawbacks such as the cost of CytoDiff reagent, but this can be balanced by the reduction in technician time required for manual counting and the use of a standardized product. The flow cytometry appears as a technique completely adapted to provide a WBC differential on a cord blood sample. The additional cell subset counts offered in the same turn-around-time would probably be exploited in the near future to better characterize the engraftment potential. The study was published in the February 2013 issue of the International Journal of Laboratory Hematology.

Related Links:
Centre Hospitalier Universitaire
Beckman Coulter



VIRCELL
DiagCor Bioscience
ADVANCED INSTRUMENTS
comments powered by Disqus
Life Technologies

Channels

Genetic Tests

view channel

Next Generation Sequencing Reveals Genetic Basis for Congenital Cataracts

Applying the latest high-throughput, next-generation DNA sequencing technology has enabled researchers in the United Kingdom to diagnose children with rare diseases that are characterized by the formation of congenital cataracts. Diagnosing which of the more than 100 diseases that cause between 20,000 and 40,000 cases... Read more

Immunology

view channel
Image: The fluorescence-activated cell sorting FACSCanto II flow cytometer (Photo courtesy of BD Biosystems).

Flow Cytometry Detects Lymphoproliferative Disorders in Fluid Specimens

Immunophenotypic analysis of hematopoietic cell populations by flow cytometry has emerged as a useful ancillary study in the diagnostic evaluation of serous effusions and cerebrospinal fluids (CSFs).... Read more

Pathology

view channel
Image: Histopathology of increased endomysial connective tissue from a patient with Duchenne Muscular Dystrophy. Necrotic fibers arrowed (Photo courtesy of Washington University in St. Louis).

New Partnership Designed to Automate Dystrophin Measurement

A developer of innovative ribonucleic acid (RNA)-based therapeutics, Sarepta Therapeutics, Inc. ( Cambridge, MA, USA), and a leading tissue-based companion diagnostics firm, Flagship Biosciences LLC (Boulder,... Read more

Lab Tech.

view channel
Image: A technical breakthrough for DNA imaging has been achieved that should quicken diagnosis of diseases for which analysis of DNA from single-cells is critical, such as early stage cancers and various pre-natal conditions (Photo courtesy of McGill University and Génome Québec Innovation Center).

Breakthrough DNA Analysis Technology to Hasten Problem Diagnosis

Researchers have achieved a technical breakthrough that should result in speedier diagnosis of diseases for which analysis of DNA from single-cells is critical, such as early stage cancers and various... Read more

Industry News

view channel

Most IVD Sector Companies Look Outside the EU and USA for Growth

Kalorama Information (New York City, NY, USA) has now published the 9th edition of its biennial market research report “Worldwide Market for In Vitro Diagnostics,” revealing estimates from its survey of the in vitro diagnostics (IVD) industry – presenting the trends, technologies, customer needs, and major suppliers with... Read more
 
Copyright © 2000-2014 Globetech Media. All rights reserved.