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Related Experiment Video

Updated: May 13, 2026

Single-cell Analysis of Immunophenotype and Cytokine Production in Peripheral Whole Blood via Mass Cytometry

Single-cell Analysis of Immunophenotype and Cytokine Production in Peripheral Whole Blood via Mass Cytometry

Published on: June 26, 2018

Creating a robust framework for the analysis of cryopreserved samples in quantitative immunological experiments.

B Ogunjimi¹, N Hens, R Malfait

¹Centre for Health Economics Research and Modeling Infectious Diseases, Vaccine & Infectious Disease Institute, University of Antwerp, Antwerp, Belgium. benson.ogunjimi@ua.ac.be

Journal of Immunological Methods

|March 26, 2013

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Cryopreservation impacts flow cytometry cell counts, particularly for CD62L and CD4+CD25+Foxp3+ markers. Basic lymphocyte subsets remain robust for longitudinal studies when using absolute counts.

Area of Science:

Immunology
Cell Biology
Biotechnology

Background:

Longitudinal immunological studies often require cryopreserved samples for flow cytometry.
Previous research indicated cryopreservation reduces CD62L and CD25+Foxp3+ counts.
The impact of cryopreservation on longitudinal cell type calculations is not well-established.

Purpose of the Study:

To compare the effects of cryopreservation on cell type calculations in longitudinal flow cytometry datasets.
To assess the robustness of various lymphocyte subset markers after cryopreservation.

Main Methods:

Compared cell counts from fresh samples with those cryopreserved for 5-6 months in 9 individuals.
Analyzed basic lymphocyte subpopulations (CD3+CD4+, CD3+CD8+, CD3-CD19+, CD3-CD56+) and further subtypes.

Related Experiment Videos

Last Updated: May 13, 2026

Single-cell Analysis of Immunophenotype and Cytokine Production in Peripheral Whole Blood via Mass Cytometry

Single-cell Analysis of Immunophenotype and Cytokine Production in Peripheral Whole Blood via Mass Cytometry

Published on: June 26, 2018

Performed longitudinal calculations using cell count differences and ratios between fresh and cryopreserved samples.

Main Results:

Basic lymphocyte differentiation counts (CD3+CD4+, CD3+CD8+, CD3-CD19+, CD3-CD56+) were robust after cryopreservation.
Subtyping revealed CCR7 and CD45RA had a relation between fresh and cryopreserved counts, unlike CD62L.
CD4+CD25+Foxp3+ counts were approximately halved after cryopreservation.
Absolute cell counts showed a one-to-one relation for most markers, except CD62L and CD4+CD25+Foxp3+.

Conclusions:

Cryopreservation significantly affects CD62L and CD4+CD25+Foxp3+ marker counts in longitudinal flow cytometry.
CD62L and CD4+CD25+Foxp3+ are not recommended for calculations on cryopreserved longitudinal datasets.
Basic lymphocyte markers are relatively robust for longitudinal studies when absolute counts are utilized.