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Determination of blood cell subtype concentrations from frozen whole blood samples using TruCount beads

Journal article
Authors Cecilia Langenskiöld
Karin Mellgren
Jonas Abrahamsson
Mats Bemark
Published in Cytometry Part B - Clinical Cytometry
Volume 94
Issue 4
Pages 660-666
ISSN 1552-4949
Publication year 2018
Published at Institute of Biomedicine, Department of Microbiology and Immunology
Institute of Clinical Sciences, Department of Pediatrics
Pages 660-666
Language en
Links dx.doi.org/10.1002/cyto.b.21390
Keywords Absolute cell count , Clinical trial , Cryopreservation , Flow cytometry , Immunophenotyping , Peripheral blood , TruCount
Subject categories Immunology in the medical area

Abstract

© 2016 International Clinical Cytometry Society.Background: In many studies it would be advantageous if blood samples could be collected and analyzed using flow cytometry at a later stage. Ideally, sample collection should involve little hands-on time, allow for long-term storage, and minimally influence the samples. Methods: Here we establish a flow cytometry antibody panel that can be used to determine granulocytes, monocytes, and lymphocyte subset concentrations in fresh and frozen whole blood using TruCount technology. Results: The panel can be used on fresh whole-blood samples as well as whole-blood samples that have been frozen after mixing with 10% DMSO. Concentrations in frozen and fresh sample is highly correlated both when frozen within 4 h and the day after collection (r≥0.98), and the estimated concentration in frozen samples was between 91 and 94% of that in fresh samples for all cell types. Conclusion: Using this method whole-blood samples can be frozen using a simple preparation method, and stored long-term before accurate determination of cell concentration. This allows for standardized analysis of the samples at a reference laboratory in multi-center studies.

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