A comprehensive assessment of four whole blood stabilisers for flow-cytometric analysis of leukocyte populations

Published: 11 July 2022| Version 1 | DOI: 10.17632/5j95wxc69v.1
Contributors:
, Ngoc Anh Nguyen

Description

We evaluated and compared head-to-head the performance of four commercial whole blood cryopreservation kits for conventional flow cytometry; 1) Cytodelics, 2) Stable-Lyse V2 and Stable-Store V2 (SLSS-V2), 3) Proteomic stabiliser (PROT-1), and 4) Transfix. We compared the stabilizers' performance to BD FACS lysing solution - treated samples and/or isolated PBMCs using the identical staining panels (table: flow_panels) and instrument settings. We used three panels to evaluate how well cryopreservation maintains the proportions of main leucocyte lineages (immune cell lineage), how well granulocyte activation markers are preserved after activation (with N-Formylmethionyl-leucyl-phenylalanine and lipopolysaccharides) (granulocyte stimulation data), and how well main T cell maturation stages can be studied after whole blood cryopreservation (T cell data). Anonymous healthy donors were used for the testing of the stabilizers. Donors (DN): n=10, age mean 29 [range 22 - 40], female 6 (60%) / male 4 (40%). Data was generated by Ngoc Anh Nguyen at Translational Immunology Research Program, University of Helsinki in years 2020-22.

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Steps to reproduce

Samples were treated with the indicated cryopreservation kits according to the manufacturers' instructions and stained for flow cytometry. Antibodies used are listed in the attached table. Data was acquired with BD LSRIII Fortessa. Data contains the compensation files used for particular analysis, we used compensation beads for the single-stained compensation controls. Data files contain the information about used stimuli.

Institutions

Helsingin Yliopisto

Categories

Cryopreservation, Flow Cytometry

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