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

Cytometry B Clin Cytom. 2018 Jul;94(4):660-666. doi: 10.1002/cyto.b.21390. Epub 2016 Jul 28.

Abstract

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. © 2016 International Clinical Cytometry Society.

Keywords: TruCount; absolute cell count; clinical trial; cryopreservation; flow cytometry; immunophenotyping; peripheral blood.

MeSH terms

  • Adult
  • Aged
  • Cryopreservation*
  • Female
  • Flow Cytometry / methods*
  • Humans
  • Immunophenotyping / methods*
  • Leukocyte Count / methods*
  • Male
  • Middle Aged
  • Specimen Handling
  • Young Adult