Journal
CYTOMETRY PART B-CLINICAL CYTOMETRY
Volume 94, Issue 4, Pages 660-666Publisher
WILEY
DOI: 10.1002/cyto.b.21390
Keywords
cryopreservation; TruCount; absolute cell count; clinical trial; flow cytometry; immunophenotyping; peripheral blood
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BackgroundIn 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. MethodsHere 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. ResultsThe 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 (r0.98), and the estimated concentration in frozen samples was between 91 and 94% of that in fresh samples for all cell types. ConclusionUsing 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. (c) 2016 International Clinical Cytometry Society
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